Heteroaryls and uses thereof

ABSTRACT

This invention provides compounds of formula I: 
     
       
         
         
             
             
         
       
         
         
           
             wherein R 1 , R 2 , CY, Y 1 , Y 2 , X 1 , X 2 , and X 3  are as described in the specification. The compounds are inhibitors of PI3K and are thus useful for treating proliferative, inflammatory, or cardiovascular disorders.

This application claims priority to U.S. provisional application No.61/132,484, filed Jun. 19, 2008, the contents of which are incorporatedby reference herein.

BACKGROUND OF THE INVENTION

Phosphatidylinositol 3-kinase (PI3K) is a family of lipid kinases thatphosphorylate phosphatidylinositol at the 3′ position of the inositolring. PI3K is comprised of several classes of genes, including Class IA,IB, II and III and some of these classes contain several isoforms(reviewed in Engelman et al., Nature Review Genetics 7:606-619 (2006)).Adding to the complexity of this family is the fact that PI3Ks functionas heterodimers, comprising a catalytic domain and a regulatory domain.The PI3K family is structurally related to a larger group of lipid andserine/threonine protein kinases known as the phosphatidylinositol3-kinase like kinases (PIKKs), which also includes DNA-PK, ATM, ATR,mTOR, TRRAP and SMG1.

PI3K is activated downstream of various mitogenic signals mediatedthrough receptor tyrosine kinases, and subsequently stimulates a varietyof biological outcomes; including increased cell survival, cell cycleprogression, cell growth, cell metabolism, cell migration andangiogenesis (reviewed in Cantley, Science 296:1655-57 (2002); Hennessyet al., Nature Reviews Drug Discovery 4:988-1004 (2005); Engelman etal., Nature Review Genetics 7:606-619 (2006)). Thus, PI3Khyper-activation is associated with a number of hyper-proliferative,inflammatory, or cardiovascular disorders; including cancer,inflammation, and cardiovascular disease.

There are a number of genetic aberrations that lead to constitutive PI3Ksignaling; including activating mutations in PI3K itself (Hennessy etal., Nature Reviews Drug Discovery 4:988-1004 (2005); reviewed in Baderet al., Nature Reviews Cancer 5:921-9 (2005)); RAS (reviewed in DownwardNature Reviews Cancer 3:11-22 (2003)) and upstream receptor tyrosinekinases (reviewed in Zwick et al., Trends in Molecular Medicine 8:17-23(2002)) as well as inactivating mutations in the tumor suppressor PTEN(reviewed in Cully et al., Nature Reviews Cancer 6:184-92 (2006)).Mutations in each of these gene classes have proven to be oncogenic andare commonly found in a variety of cancers.

The molecules defined within this invention inhibit the activity ofPI3K, and therefore may be useful for the treatment of proliferative,inflammatory, or cardiovascular disorders. Cases where PI3K pathwaymutations have been linked to proliferative disorders where themolecules defined within this invention may have a therapeutic benefitinclude benign and malignant tumors and cancers from diverse lineage,including but not limited to those derived from colon (Samuels et al.,Science 304:554 (2004); reviewed in Karakas et al., British Journal ofCancer 94: 455-59 (2006)), liver (reviewed in Karakas et al., BritishJournal of Cancer 94: 455-59 (2006)), intestine (reviewed in Hennessy etal., Nature Reviews Drug Discovery 4:988-1004 (2005)), stomach (Samuelset al., Science 304:554 (2004); reviewed in Karakas et al., BritishJournal of Cancer 94: 455-59 (2006)), esophagus (Phillips et al.,International Journal of Cancer 118:2644-6 (2006)); pancreas (reviewedin Downward Nature Reviews Cancer 3:11-22 (2003)); skin (reviewed inHennessy et al., Nature Reviews Drug Discovery 4:988-1004 (2005)),prostate (reviewed in Hennessy et al., Nature Reviews Drug Discovery4:988-1004 (2005)), lung (Samuels et al., Science 304:554 (2004);reviewed in Karakas et al., British Journal of Cancer 94: 455-59(2006)), breast (Samuels et al., Science 304:554 (2004); Isakoff et al.,Can Res 65:10992-1000 (2005); reviewed in Karakas et al., BritishJournal of Cancer 94: 455-59 (2006)), endometrium (Oda et al., Can Res65:10669-73 (2005); reviewed in Hennessy et al., Nature Reviews DrugDiscovery 4:988-1004 (2005)), cervix (reviewed in Hennessy et al.,Nature Reviews Drug Discovery 4:988-1004 (2005)); ovary (Shayesteh etal., Nature Genetics 21:99-102 (1999); reviewed in Karakas et al.,British Journal of Cancer 94: 455-59 (2006)), testes (Moul et al., GenesChromosomes Cancer 5:109-18 (1992); Di Vizio et al., Oncogene 24:1882-94(2005)), hematological cells (reviewed in Karakas et al., BritishJournal of Cancer 94: 455-59 (2006); Hennessy et al., Nature ReviewsDrug Discovery 4:988-1004 (2005)), pancreas (reviewed in Downward NatureReviews Cancer 3:11-22 (2003)), thyroid (reviewed in Downward NatureReviews Cancer 3:11-22 (2003); reviewed in Hennessy et al., NatureReviews Drug Discovery 4:988-1004 (2005)); brain (Samuels et al.,Science 304:554 (2004); reviewed in Karakas et al., British Journal ofCancer 94: 455-59 (2006)), bladder (Lopez-Knowles et al., CancerResearch 66:7401-7404 (2006); Hennessy et al., Nature Reviews DrugDiscovery 4:988-1004 (2005)); kidney (reviewed in Downward NatureReviews Cancer 3:11-22 (2003)) and Head and Neck (reviewed in Engelmanet al., Nature Reviews Genetics 7:606-619 (2006)).

Other classes of disorders with aberrant PI3K pathway signaling wherethe molecules defined within this invention may have a therapeuticbenefit include inflammatory and cardiovascular diseases, including butnot limited to allergies/anaphylaxis (reviewed in Rommel et al., NatureReviews Immunology 7:191-201 (2007)), acute and chronic inflammation(reviewed in Ruckle et al., Nature Reviews Drug Discovery 5:903-12(2006); reviewed in Rommel et al., Nature Reviews Immunology 7:191-201(2007)), rheumatoid arthritis (reviewed in Rommel et al., Nature ReviewsImmunology 7:191-201 (2007)); autoimmunity disorders (reviewed in Ruckleet al., Nature Reviews Drug Discovery 5:903-12 (2006)), thrombosis(Jackson et al., Nature Medicine 11:507-14 (2005); reviewed in Ruckle etal., Nature Reviews Drug Discovery 5:903-12 (2006)), hypertension(reviewed in Ruckle et al., Nature Reviews Drug Discovery 5:903-12(2006)), cardiac hypertrophy (reviewed in Proud et al., CardiovascularResearch 63:403-13 (2004)), and heart failure (reviewed in Mocanu etal., British Journal of Pharmacology 150:833-8 (2007)).

Clearly, it would be beneficial to provide novel PI3K inhibitors thatpossess good therapeutic properties, especially for the treatment ofproliferative, inflammatory, or cardiovascular disorders.

DETAILED DESCRIPTION OF THE INVENTION 1. General Description ofCompounds of the Invention

This invention provides compounds that are inhibitors of PI3K, andaccordingly are useful for the treatment of proliferative, inflammatory,or cardiovascular disorders. The compounds of this invention arerepresented by formula I:

or a pharmaceutically acceptable salt thereof, wherein:

when Y₂ is C, R¹ is H, —CN, halogen, —Z—R³, C₁₋₆ aliphatic, or3-10-membered cycloaliphatic, wherein:

-   -   Z is selected from an optionally substituted C₁₋₃ alkylene        chain, —O—, —N(R^(1a))—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —CO₂—,        —C(O)NR^(1a)—, —N(R^(1a))C(O)—, —N(R^(1a))CO₂—, —S(O)₂NR^(1a)—,        —N(R^(1a))S(O)₂—, —OC(O)N(R^(1a))—, —N(R^(1a))C(O)NR^(1a)—,        —N(R^(1a))S(O)₂N(R^(1a))—, or —OC(O)—;    -   R^(1a) is hydrogen or an optionally substituted C₁₋₄ aliphatic,        and    -   R³ is an optionally substituted group selected from C₁₋₆        aliphatic, 3-10-membered cycloaliphatic, 4-10-membered        heterocyclyl having 1-5 heteroatoms independently selected from        nitrogen, oxygen, or sulfur, 6-10-membered aryl, or        5-10-membered heteroaryl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur;

R² is H, halogen, —W—R⁵, or —R⁵, wherein:

-   -   W is selected from an optionally substituted C₁₋₃ alkylene        chain, —O—, —N(R^(2a))—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —CO₂—,        —C(O)NR^(2a)—, —N(R^(2a))C(O)—, —N(R^(2a))CO₂—, —S(O)₂NR^(2a)—,        —N(R^(2a))S(O)₂—, —OC(O)N(R^(2a))—, —N(R^(2a))C(O)NR^(2a)—,        —N(R^(2a))S(O)₂N(R^(2a))—, or —OC(O)—.    -   R^(2a) is hydrogen or an optionally substituted C₁₋₄ aliphatic,        and    -   R⁵ is an optionally substituted group selected from C₁₋₆        aliphatic, 3-10-membered cycloaliphatic, 4-10-membered        heterocyclyl having 1-5 heteroatoms independently selected from        nitrogen, oxygen, or sulfur, 6-10-membered aryl, or        5-10-membered heteroaryl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur;        X₁, X₂, and X₃ are each independently N or CR⁶, wherein each        occurrence of R⁶ is independently hydrogen, —CN, halogen, —V—R⁷,        C₁₋₆ aliphatic, or 3-10-membered cycloaliphatic, wherein:    -   V is selected from an optionally substituted C₁₋₃ alkylene        chain, —O—, —N(R^(6a))—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —CO₂—,        —C(O)NR^(6a)—, —N(R^(6a))C(O)—, —N(R^(6a))CO₂—, —S(O)₂NR^(6a)—,        —N(R^(6a))S(O)₂—, —OC(O)N(R^(6a))—, —N(R^(6a))C(O)NR^(6a)—,        —N(R^(6a))S(O)₂N(R^(6a))—, or —OC(O)—.    -   R^(6a) is hydrogen or an optionally substituted C₁₋₄ aliphatic,        and    -   R⁷ is an optionally substituted group selected from C₁₋₆        aliphatic, 3-10-membered cycloaliphatic, 4-10-membered        heterocyclyl having 1-5 heteroatoms independently selected from        nitrogen, oxygen, or sulfur, 6-10-membered aryl, or        5-10-membered heteroaryl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur;    -   Y₁ is S, O, NR⁸, wherein R⁸ is hydrogen or an optionally        substituted C₁₋₄aliphatic;    -   Y₂ is C, N

CY is

wherein each occurrence of R⁴ is independently-R^(4a) or -T₁-R^(4d),wherein:

-   -   each occurrence of R^(4a), as valency and stability permit, is        independently fluorine, ═O, ═S, —CN, —NO₂, —R^(4c), N(R^(4b))₂,        —OR^(4b), —SR^(4c), —S(O)₂R^(4c), —C(O)R^(4b), —C(O)OR^(4b),        —C(O)N(R^(4b))₂, —S(O)₂N(R^(4b))₂, —OC(O)N(R^(4b))₂,        —N(R^(4e))C(O)R^(4b), —N(R^(4e))SO₂R^(4c),        —N(R^(4e))C(O)OR^(4b), —N(R^(4e))C(O)N(R^(4b))₂, or        —N(R^(4e))SO₂N(R^(4b))₂, or two occurrences of R^(4b), taken        together with a nitrogen atom to which they are bound, form an        optionally substituted 4-7-membered heterocyclyl ring having 0-1        additional heteroatoms selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(4b) is independently hydrogen or an        optionally substituted group selected from C₁-C₆ aliphatic,        3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(4c) is independently an optionally        substituted group selected from C₁-C₆ aliphatic, 3-10-membered        cycloaliphatic, 4-10-membered heterocyclyl having 1-5        heteroatoms independently selected from nitrogen, oxygen, or        sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(4d) is independently hydrogen or an        optionally substituted from 3-10-membered cycloaliphatic,        4-10-membered heterocyclyl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl,        or 5-10-membered heteroaryl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur;    -   each occurrence of R^(4e) is independently hydrogen or an        optionally substituted C₁₋₆ aliphatic group; and    -   T₁ is an optionally substituted C₁-C₆ alkylene chain wherein the        alkylene chain optionally is interrupted by —N(R^(4a))—, —O—,        —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(4a))—,        —S(O)₂N(R^(4a))—, —OC(O)N(R^(4a))—, —N(R^(4a))C(O)—,        —N(R^(4a))SO₂—, N(R^(4a))C(O)O—, —NR^(4a)C(O)N(R^(4a))—,        —N(R^(4a))S(O)₂N(R^(4a))—, —OC(O)—, or —C(O)N(R^(4a))—O— or        wherein T₁ or a portion thereof optionally forms part of an        optionally substituted 3-7 membered cycloaliphatic or        heterocyclyl ring;

n is 0-6;

m is 1 or 2;

p is 0, 1, or 2;

represents a single or double bond; and provided that the compound offormula I is other than Morpholine,4-[5-(4,5-diphenyl-1H-imidazol-2-yl)-2-thienyl],

2. Compounds and Definitions

Compounds of this invention include those described generally forformula I above, and are further illustrated by the classes, subclasses,and species disclosed herein. As used herein, the following definitionsshall apply unless otherwise indicated.

As described herein, compounds of the invention may be optionallysubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the invention. It will be appreciated that the phrase“optionally substituted” is used interchangeably with the phrase“substituted or unsubstituted.” In general, the term “substituted”,whether preceded by the “optionally” or not, means that a hydrogenradical of the designated moiety is replaced with the radical of aspecified substituent, provided that the substitution results in astable or chemically feasible compound. The term “substitutable”, whenused in reference to a designated atom, means that attached to the atomis a hydrogen radical, which hydrogen atom can be replaced with theradical of a suitable substituent. Unless otherwise indicated, an“optionally substituted” group may have a substituent at eachsubstitutable position of the group, and when more than one position inany given structure may be substituted with more than one substituentselected from a specified group, the substituent may be either the sameor different at every position. Combinations of substituents envisionedby this invention are preferably those that result in the formation ofstable or chemically feasible compounds.

A stable compound or chemically feasible compound is one in which thechemical structure is not substantially altered when kept at atemperature from about −80° C. to about +40°, in the absence of moistureor other chemically reactive conditions, for at least a week, or acompound which maintains its integrity long enough to be useful fortherapeutic or prophylactic administration to a patient.

The phrase “one or more substituents”, as used herein, refers to anumber of substituents that equals from one to the maximum number ofsubstituents possible based on the number of available bonding sites,provided that the above conditions of stability and chemical feasibilityare met.

As used herein, the term “independently selected” means that the same ordifferent values may be selected for multiple instances of a givenvariable in a single compound.

As used herein, “a 3-7-membered saturated, partially unsaturated, oraromatic monocyclic ring having 0-3 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur, or an 8-10-membered partiallyunsaturated, or aromatic bicyclic ring system having 0-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur” includescycloaliphatic, heterocyclic, aryl and heteroaryl rings.

As used herein, the term “aromatic” includes aryl and heteroaryl groupsas described generally below and herein.

The term “aliphatic” or “aliphatic group”, as used herein, means anoptionally substituted straight-chain or branched C₁₋₁₂ hydrocarbon, ora cyclic C₁₋₁₂ hydrocarbon which is completely saturated or whichcontains one or more units of unsaturation, but which is not aromatic(also referred to herein as “carbocycle”, “cycloaliphatic”,“cycloalkyl”, or “cycloalkenyl”). For example, suitable aliphatic groupsinclude optionally substituted linear, branched or cyclic alkyl,alkenyl, alkynyl groups and hybrids thereof, such as (cycloalkyl)alkyl,(cycloalkenyl)alkyl, or (cycloalkyl)alkenyl. Unless otherwise specified,in various embodiments, aliphatic groups have 1-12, 1-10, 1-8, 1-6, 1-4,1-3, or 1-2 carbon atoms.

The term “alkyl”, used alone or as part of a larger moiety, refers to anoptionally substituted straight or branched chain hydrocarbon grouphaving 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms.

The term “alkenyl”, used alone or as part of a larger moiety, refers toan optionally substituted straight or branched chain hydrocarbon grouphaving at least one double bond and having 2-12, 2-10, 2-8, 2-6, 2-4, or2-3 carbon atoms.

The term “alkynyl”, used alone or as part of a larger moiety, refers toan optionally substituted straight or branched chain hydrocarbon grouphaving at least one triple bond and having 2-12, 2-10, 2-8, 2-6, 2-4, or2-3 carbon atoms.

The terms “cycloaliphatic”, “carbocycle”, “carbocyclyl”, “carbocyclo”,or “carbocyclic”, used alone or as part of a larger moiety, refer to anoptionally substituted saturated or partially unsaturated cyclicaliphatic ring system having from 3 to about 14 ring carbon atoms. Insome embodiments, the cycloaliphatic group is an optionally substitutedmonocyclic hydrocarbon having 3-8 or 3-6 ring carbon atoms.Cycloaliphatic groups include, without limitation, optionallysubstituted cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl,cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl,cyclooctenyl, or cyclooctadienyl. The terms “cycloaliphatic”,“carbocycle”, “carbocyclyl”, “carbocyclo”, or “carbocyclic” also includeoptionally substituted bridged or fused bicyclic rings having 6-12,6-10, or 6-8 ring carbon atoms, wherein any individual ring in thebicyclic system has 3-8 ring carbon atoms.

The term “cycloalkyl” refers to an optionally substituted saturated ringsystem of about 3 to about 10 ring carbon atoms. Exemplary monocycliccycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, and cycloheptyl.

The term “cycloalkenyl” refers to an optionally substituted non-aromaticmonocyclic or multicyclic ring system containing at least onecarbon-carbon double bond and having about 3 to about 10 carbon atoms.Exemplary monocyclic cycloalkenyl rings include cyclopentyl,cyclohexenyl, and cycloheptenyl.

The terms “haloaliphatic”, “haloalkyl”, “haloalkenyl” and “haloalkoxy”refer to an aliphatic, alkyl, alkenyl or alkoxy group, as the case maybe, which is substituted with one or more halogen atoms. As used herein,the term “halogen” or “halo” means F, Cl, Br, or I. The term“fluoroaliphatic” refers to a haloaliphatic wherein the halogen isfluoro, including perfluorinated aliphatic groups. Examples offluoroaliphatic groups include, without limitation, fluoromethyl,difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl,1,1,2-trifluoroethyl, 1,2,2-trifluoroethyl, and pentafluoroethyl.

The term “heteroatom” refers to one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The tennis “aryl” and “ar-”, used alone or as part of a larger moiety,e.g., “aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refer to an optionallysubstituted C₆₋₁₄aromatic hydrocarbon moiety comprising one to threearomatic rings. Preferably, the aryl group is a C₆₋₁₀aryl group. Arylgroups include, without limitation, optionally substituted phenyl,naphthyl, or anthracenyl. The teens “aryl” and “ar-”, as used herein,also include groups in which an aryl ring is fused to one or morecycloaliphatic rings to form an optionally substituted cyclic structuresuch as a tetrahydronaphthyl, indenyl, or indenyl ring. The term “aryl”may be used interchangeably with the terms “aryl group”, “aryl ring”,and “aromatic ring”.

An “aralkyl” or “arylalkyl” group comprises an aryl group covalentlyattached to an alkyl group, either of which independently is optionallysubstituted. Preferably, the aralkyl group is C₆₋₁₀ arylC₁₋₆alkyl,including, without limitation, benzyl, phenethyl, and naphthylmethyl.

The terms “heteroaryl” and “heteroar-”, used alone or as part of alarger moiety, e.g., “heteroaralkyl”, or “heteroaralkoxy”, refer togroups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms;having 6, 10, or 14 π electrons shared in a cyclic array; and having, inaddition to carbon atoms, from one to five heteroatoms. A heteroarylgroup may be mono-, bi-, tri-, or polycyclic, preferably mono-, bi-, ortricyclic, more preferably mono- or bicyclic. The term “heteroatom”refers to nitrogen, oxygen, or sulfur, and includes any oxidized form ofnitrogen or sulfur, and any quaternized form of a basic nitrogen. Forexample, a nitrogen atom of a heteroaryl may be a basic nitrogen atomand may also be optionally oxidized to the corresponding N-oxide. When aheteroaryl is substituted by a hydroxy group, it also includes itscorresponding tautomer. The terms “heteroaryl” and “heteroar-”, as usedherein, also include groups in which a heteroaromatic ring is fused toone or more aryl, cycloaliphatic, or heterocycloaliphatic rings.Nonlimiting examples of heteroaryl groups include thienyl, furanyl,pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl,isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl,pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl,naphthyridinyl, pteridinyl, indolyl, isoindolyl, benzothienyl,benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl,quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl,quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl,phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Theterm “heteroaryl” may be used interchangeably with the terms “heteroarylring”, “heteroaryl group”, or “heteroaromatic”, any of which termsinclude rings that are optionally substituted. The term “heteroaralkyl”refers to an alkyl group substituted by a heteroaryl, wherein the alkyland heteroaryl portions independently are optionally substituted.

As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclicradical”, and “heterocyclic ring” are used interchangeably and refer toa stable 3- to 8-membered monocyclic or 7-10-membered bicyclicheterocyclic moiety that is either saturated or partially unsaturated,and having, in addition to carbon atoms, one or more, preferably one tofour, heteroatoms, as defined above. When used in reference to a ringatom of a heterocycle, the term “nitrogen” includes a substitutednitrogen. As an example, in a saturated or partially unsaturated ringhaving 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, thenitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as inpyrrolidinyl), or NR⁺ (as in N-substituted pyrrolidinyl).

A heterocyclic ring can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted. Examples of such saturatedor partially unsaturated heterocyclic radicals include, withoutlimitation, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl,decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl,diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiamorpholinyl. Aheterocyclyl group may be mono-, bi-, tri-, or polycyclic, preferablymono-, bi-, or tricyclic, more preferably mono- or bicyclic. The term“heterocyclylalkyl” refers to an alkyl group substituted by aheterocyclyl, wherein the alkyl and heterocyclyl portions independentlyare optionally substituted. Additionally, a heterocyclic ring alsoincludes groups in which the heterocyclic ring is fused to one or morearyl rings.

As used herein, the term “partially unsaturated” refers to a ring moietythat includes at least one double or triple bond between ring atoms. Theterm “partially unsaturated” is intended to encompass rings havingmultiple sites of unsaturation, but is not intended to include aromatic(e.g., aryl or heteroaryl) moieties, as herein defined.

The term “alkylene” refers to a bivalent alkyl group. An “alkylenechain” is a polymethylene group, i.e., —(CH₂)_(n)—, wherein n is apositive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from1 to 2, or from 2 to 3. An optionally substituted alkylene chain is apolymethylene group in which one or more methylene hydrogen atoms isoptionally replaced with a substituent. Suitable substituents includethose described below for a substituted aliphatic group and also includethose described in the specification herein. It will be appreciated thattwo substituents of the alkylene group may be taken together to form aring system. In certain embodiments, two substituents can be takentogether to form a 3-7-membered ring. The substituents can be on thesame or different atoms.

An alkylene chain also can be optionally interrupted by a functionalgroup. An alkylene chain is “interrupted” by a functional group when aninternal methylene unit is interrupted by the functional group. Examplesof suitable “interrupting functional groups” are described in thespecification and claims herein.

For purposes of clarity, all bivalent groups described herein,including, e.g., the alkylene chain linkers described above, areintended to be read from left to right, with a correspondingleft-to-right reading of the formula or structure in which the variableappears.

An aryl (including aralkyl, aralkoxy, aryloxyalkyl and the like) orheteroaryl (including heteroaralkyl and heteroarylalkoxy and the like)group may contain one or more substituents and thus may be “optionallysubstituted”. In addition to the substituents defined above and herein,suitable substituents on the unsaturated carbon atom of an aryl orheteroaryl group also include and are generally selected from -halo,—NO₂, —CN, —R⁺, —C(R⁺)═C(R⁺)₂, —C≡C—R⁺, —OR⁺, —SR^(o), —S(O)R^(o),—SO₂R^(o), —SO₃R⁺, —SO₂N(R⁺)₂, —N(R⁺)₂, —NR⁺C(O)R⁺, —NR⁺C(S)R⁺,—NR⁺C(O)N(R⁺)₂, —NR⁺C(S)N(R⁺)₂, —N(R⁺)C(═NR⁺)—N(R⁺)₂,—N(R⁺)C(═NR⁺)—R^(o), —NR⁺CO₂R⁺, —NR⁺SO₂R^(o), —NR⁺SO₂N(R⁺)₂, —O—C(O)R⁺,—O—CO₂R⁺, —OC(O)N(R⁺)₂, —C(O)R⁺, —C(S)R^(o), —CO₂R⁺, —C(O)—C(O)R⁺,—C(O)N(R⁺)₂, —C(S)N(R⁺)₂, —C(O)N(R⁺)—OR⁺, —C(O)N(R⁺)C(═NR⁺)—N(R⁺)₂,—N(R⁺)C(═NR⁺)—N(R⁺)—C(O)R⁺, —C(═NR⁺)—N(R⁺)₂, —C(═NR⁺)—OR⁺,—N(R⁺)—N(R⁺)₂, —C(═NR⁺)—N(R⁺)—OR⁺, —C(R^(o))═N—OR⁺, —P(O)(R⁺)₂,—P(O)(OR⁺)₂, —O—P(O)—OR⁺, and —P(O)(NR⁺)—N(R⁺)₂, wherein R⁺,independently, is hydrogen or an optionally substituted aliphatic, aryl,heteroaryl, cycloaliphatic, or heterocyclyl group, or two independentoccurrences of R⁺ are taken together with their intervening atom(s) toform an optionally substituted 5-7-membered aryl, heteroaryl,cycloaliphatic, or heterocyclyl ring. Each R^(o) is an optionallysubstituted aliphatic, aryl, heteroaryl, cycloaliphatic, or heterocyclylgroup.

An aliphatic or heteroaliphatic group, or a non-aromatic carbycyclic orheterocyclic ring may contain one or more substituents and thus may be“optionally substituted”. Unless otherwise defined above and herein,suitable substituents on the saturated carbon of an aliphatic orheteroaliphatic group, or of a non-aromatic carbocyclic or heterocyclicring are selected from those listed above for the unsaturated carbon ofan aryl or heteroaryl group and additionally include the following: ═O,═S, ═C(R*)₂, ═N—N(R*)₂, ═N—OR*, ═N—NHC(O)R*, ═N—NHCO₂R^(o)═N—NHSO₂R^(o)or ═N—R* where R^(o) is defined above, and each R* is independentlyselected from hydrogen or an optionally substituted C₁₋₆ aliphaticgroup.

In addition to the substituents defined above and herein, optionalsubstituents on the nitrogen of a non-aromatic heterocyclic ring alsoinclude and are generally selected from —R⁺, —N(R⁺)₂, —C(O)R⁺, —C(O)OR⁺,—C(O)C(O)R⁺, —C(O)CH₂C(O)R⁺, —S(O)₂R⁺, —S(O)₂N(R⁺)₂, —C(S)N(R⁺)₂,—C(═NH)—N(R⁺)₂, or —N(R⁺)S(O)₂R⁺; wherein each R⁺ is defined above. Aring nitrogen atom of a heteroaryl or non-aromatic heterocyclic ringalso may be oxidized to form the corresponding N-hydroxy or N-oxidecompound. A nonlimiting example of such a heteroaryl having an oxidizedring nitrogen atom is N-oxidopyridyl.

As detailed above, in some embodiments, two independent occurrences ofR⁺ (or any other variable similarly defined in the specification andclaims herein), are taken together with their intervening atom(s) toform a monocyclic or bicyclic ring selected from 3-13-memberedcycloaliphatic, 3-12-membered heterocyclyl having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, 6-10-memberedaryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur.

Exemplary rings that are formed when two independent occurrences of R⁺(or any other variable similarly defined in the specification and claimsherein), are taken together with their intervening atom(s) include, butare not limited to the following: a) two independent occurrences of R⁺(or any other variable similarly defined in the specification or claimsherein) that are bound to the same atom and are taken together with thatatom to form a ring, for example, N(R⁺)₂, where both occurrences of R⁺are taken together with the nitrogen atom to form a piperidin-1-yl,piperazin-1-yl, or morpholin-4-yl group; and b) two independentoccurrences of R⁺ (or any other variable similarly defined in thespecification or claims herein) that are bound to different atoms andare taken together with both of those atoms to form a ring, for examplewhere a phenyl group is substituted with two occurrences of OR⁺

these two occurrences of R⁺ are taken together with the oxygen atoms towhich they are bound to form a fused 6-membered oxygen containing ring:

It will be appreciated that a variety of other rings (e.g., Spiro andbridged rings) can be formed when two independent occurrences of R⁺ (orany other variable similarly defined in the specification and claimsherein) are taken together with their intervening atom(s) and that theexamples detailed above are not intended to be limiting.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention. Unless otherwise stated, alltautomeric forms of the compounds of the invention are within the scopeof the invention. Additionally, unless otherwise stated, structuresdepicted herein are also meant to include compounds that differ only inthe presence of one or more isotopically enriched atoms. For example,compounds having the present structures except for the replacement ofhydrogen by deuterium or tritium, or the replacement of a carbon by a¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. Suchcompounds are useful, for example, as analytical tools or probes inbiological assays.

It is to be understood that, when a disclosed compound has at least onechiral center, the present invention encompasses one enantiomer ofinhibitor free from the corresponding optical isomer, racemic mixture ofthe inhibitor and mixtures enriched in one enantiomer relative to itscorresponding optical isomer. When a mixture is enriched in oneenantiomer relative to its optical isomers, the mixture contains, forexample, an enantiomeric excess of at least 50%, 75%, 90%, 95% 99% or99.5%.

The enantiomers of the present invention may be resolved by methodsknown to those skilled in the art, for example by formation ofdiastereoisomeric salts which may be separated, for example, bycrystallization; formation of diastereoisomeric derivatives or complexeswhich may be separated, for example, by crystallization, gas-liquid orliquid chromatography; selective reaction of one enantiomer with anenantiomer-specific reagent, for example enzymatic esterification; orgas-liquid or liquid chromatography in a chiral environment, for exampleon a chiral support for example silica with a bound chiral ligand or inthe presence of a chiral solvent. Where the desired enantiomer isconverted into another chemical entity by one of the separationprocedures described above, a further step is required to liberate thedesired enantiomeric form. Alternatively, specific enantiomers may besynthesized by asymmetric synthesis using optically active reagents,substrates, catalysts or solvents, or by converting one enantiomer intothe other by asymmetric transformation.

When a disclosed compound has at least two chiral centers, the presentinvention encompasses a diastereomer free of other diastereomers, a pairof diastereomers free from other diasteromeric pairs, mixtures ofdiasteromers, mixtures of diasteromeric pairs, mixtures of diasteromersin which one diastereomer is enriched relative to the otherdiastereomer(s) and mixtures of diasteromeric pairs in which onediastereomeric pair is enriched relative to the other diastereomericpair(s). When a mixture is enriched in one diastereomer ordiastereomeric pair(s) relative to the other diastereomers ordiastereomeric pair(s), the mixture is enriched with the depicted orreferenced diastereomer or diastereomeric pair(s) relative to otherdiastereomers or diastereomeric pair(s) for the compound, for example,by a molar excess of at least 50%, 75%, 90%, 95%, 99% or 99.5%.

The diastereoisomeric pairs may be separated by methods known to thoseskilled in the art, for example chromatography or crystallization andthe individual enantiomers within each pair may be separated asdescribed above. Specific procedures for chromatographically separatingdiastereomeric pairs of precursors used in the preparation of compoundsdisclosed herein are provided the examples herein.

3. Description of Exemplary Compounds

In certain embodiments, for compounds of general formula I, one or moresubstituents are selected from:

(a) Y₁ is S;

(b) Y₂ is C

(c) R¹ is CN or H;

(d) R² is an optionally substituted 6-10-membered aryl, or 5-10-memberedheteroaryl having 1-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur;

(e) n is 0-2; or

(f) R⁴ is —R^(4a).

In some embodiments, a compound of formula I is represented by:

In yet other embodiments, R² is a 3-10-membered cycloaliphatic,4-10-membered heterocyclyl having 1-5 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur 6-10-membered aryl, or 5-10-memberedheteroaryl having 1-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur, optionally substituted with 1-4 independentoccurrences of R⁹, wherein R⁹ is —R^(9a), -T₂-R^(9d), or —V₂-T₂-R^(9d),and:

-   -   each occurrence of R^(9a) is independently halogen, —CN, —NO₂,        —R^(9c), —N(R^(9b))₂, —OR^(9b), —SR^(9c), —S(O)₂R^(9c),        —C(O)R^(9b), —C(O)OR^(9b), —C(O)N(R^(9b))₂, —S(O)₂N(R^(9b))₂,        —OC(O)N(R^(9b))₂, —N(R^(9e))C(O)R^(9b), —N(R^(9e))SO₂R^(9c),        —N(R^(9e))C(O)OR^(9b), —N(R^(9e))C(O)N(R^(9b))₂, or        —N(R^(9e))SO₂N(R^(9b))₂, or two occurrences of R^(9b), taken        together with a nitrogen atom to which they are bound, form an        optionally substituted 4-7-membered heterocyclyl ring having 0-1        additional heteroatoms selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(9b) is independently hydrogen or an        optionally substituted group selected from C₁-C₆ aliphatic,        3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(9c) is independently an optionally        substituted group selected from C₁-C₆aliphatic, 3-10-membered        cycloaliphatic, 4-10-membered heterocyclyl having 1-5        heteroatoms independently selected from nitrogen, oxygen, or        sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(9d) is independently hydrogen or an        optionally substituted from 3-10-membered cycloaliphatic,        4-10-membered heterocyclyl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl,        or 5-10-membered heteroaryl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur;    -   each occurrence of R^(9e) is independently hydrogen or an        optionally substituted C₁₋₆ aliphatic group;    -   each occurrence of V₂ is independently —N(R^(9e))—, —O—, —S—,        —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(9e))—,        —S(O)₂N(R^(9e))—, —OC(O)N(R^(9e))—, —N(R^(9e))C(O)—,        —N(R^(9e))SO₂—, —N(R^(9e))C(O)O—, —NR^(9e)C(O)N(R^(9e))—,        —N(R^(9e))SO₂N(R^(9e))—, —OC(O)—, or —C(O)N(R^(9e))—O—; and    -   T₂ is an optionally substituted C₁-C₆alkylene chain wherein the        alkylene chain optionally is interrupted by —N(R^(7a))—, —O—,        —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(7a))—,        —S(O)₂N(R^(7a))—, —OC(O)N(R^(7a))—, —N(R^(7a))C(O)—,        —N(R^(7a))SO₂—, —N(R^(7a))C(O)O—, —NR^(7a)C(O)N(R^(7a))—,        —N(R^(7a))S(O)₂N(R^(7a))—, —OC(O)—, or —C(O)N(R^(7a))—O— or        wherein T₃ or a portion thereof optionally forms part of an        optionally substituted 3-7 membered cycloaliphatic or        heterocyclyl ring.

In still other embodiments, Y₁ is S, Y₂ is C and the compound isrepresented by formula II:

wherein R¹ is CN or H.

In yet other embodiments, R² is an optionally substituted 3-10-memberedcycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, 6-10-memberedaryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur;

n is 0-2; and

R⁴ is —R^(4a).

In still other embodiments, a compound of formula I is represented by:

In still other embodiments, for compounds described directly above, R²is a 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5heteroatoms independently selected from nitrogen, oxygen, or sulfur6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, optionallysubstituted with 1-4 independent occurrences of R⁹, wherein R⁹ is—R^(9e), -T₂-R^(9d), or —V₂-T₂-R^(9d), and:

-   -   each occurrence of R^(9a) is independently halogen, —CN, —NO₂,        —R^(9c), —N(R^(9b))₂, —OR^(9b), —SR^(9c), —S(O)₂R^(9c),        —C(O)R^(9b), —C(O)OR^(9b), —C(O)N(R^(9b))₂, —S(O)₂N(R^(9b))₂,        —OC(O)N(R^(9b))₂, —N(R^(9e))C(O)R^(9b), —N(R^(9e))SO₂R^(9c),        —N(R^(9e))C(O)OR^(9b), —N(R^(9e))C(O)N(R^(9b))₂, or        —N(R^(9e))SO₂N(R^(9b))₂, or two occurrences of R^(9b), taken        together with a nitrogen atom to which they are bound, form an        optionally substituted 4-7-membered heterocyclyl ring having 0-1        additional heteroatoms selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(9b) is independently hydrogen or an        optionally substituted group selected from C₁-C₆ aliphatic,        3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(9c) is independently an optionally        substituted group selected from C₁-C₆ aliphatic, 3-10-membered        cycloaliphatic, 4-10-membered heterocyclyl having 1-5        heteroatoms independently selected from nitrogen, oxygen, or        sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having        1-5 heteroatoms independently selected from nitrogen, oxygen, or        sulfur;    -   each occurrence of R^(9d) is independently hydrogen or an        optionally substituted from 3-10-membered cycloaliphatic,        4-10-membered heterocyclyl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl,        or 5-10-membered heteroaryl having 1-5 heteroatoms independently        selected from nitrogen, oxygen, or sulfur;    -   each occurrence of R^(9e) is independently hydrogen or an        optionally substituted C₁₋₆ aliphatic group;    -   each occurrence of V₂ is independently —N(R^(9e))—, —O—, —S—,        —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(9e))—,        —S(O)₂N(R^(9e))—, —OC(O)N(R^(9e))—, —N(R^(9e))C(O)—,        —N(R^(9e))SO₂—, —N(R^(9e))C(O)O—, —NR^(9e)C(O)N(R^(9e))—,        —N(R^(9e))SO₂N(R^(9e))—, —OC(O)—, or —C(O)N(R^(9e))—O—; and    -   T₂ is an optionally substituted C₁-C₆ alkylene chain wherein the        alkylene chain optionally is interrupted by —N(R^(7a))—, —O—,        —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(7a))—,        —S(O)₂N(R^(7a))—, —OC(O)N(R^(7a))—, —N(R^(7a))C(O)—,        —N(R^(7a))SO₂—, —N(R^(7a))C(O)O—, —NR^(7a)C(O)N(R^(7a))—,        —N(R^(7a))S(O)₂N(R^(7a))—, —OC(O)—, or —C(O)N(R^(7a))—O— or        wherein T₃ or a portion thereof optionally forms part of an        optionally substituted 3-7 membered cycloaliphatic or        heterocyclyl ring.

In yet other embodiments, R² is a phenyl group substituted with 1-3independent occurrences of halogen, —CN, —NO₂, —R^(9c), —N(R^(9b))₂,—OR^(9b), —SR^(9c), —S(O)₂R^(9c), —C(O)R^(9b), —C(O)OR^(9b),—C(O)N(R^(9b))₂, —S(O)₂N(R^(9b))₂, —OC(O)N(R^(9b))₂,—N(R^(9e))C(O)R^(9b), —N(R^(9e))SO₂R^(9c), —N(R^(9e))C(O)OR^(9b),—N(R^(9e))C(O)N(R^(9b))₂, or —N(R^(9e))SO₂N(R^(9b))₂;

R³ is hydrogen, C₁₋₄alkyl, or C₁₋₄fluoroalkyl; and

n is 0.

In still other embodiments, R² is a phenyl group substituted with 1-3independent occurrences of halo, C₁₋₃ alkyl, CN, C₁₋₃haloalkyl, —OC₁₋₃alkyl, —OC₁₋₃haloalkyl, —NHC(O)C₁₋₃alkyl, —NHC(O)NHC₁₋₃ alkyl,NHS(O)₂C₁₋₃alkyl, or —COH.

Table 1 below depicts certain exemplary compounds of formula I:

TABLE 1 Exemplary Compounds of formula I:  

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

4 General Synthetic Methods and Intermediates

The compounds of the present invention can be prepared by methods knownto one of ordinary skill in the art and/or by reference to the schemesshown below and the synthetic examples that follow. Exemplary syntheticroutes are set forth in Schemes 1-11 below, and in the Examples.

Scheme 1 above shows a general route for preparing compounds of formula(vii). As shown in Scheme 1, conversion of amide i to thioamide ii canbe accomplished using a suitable reagent, such as Lawesson's reagent, inTHF (Method A). Thioamide ii can be coupled with acyl chlorides in thepresence of an appropriate base, such as DIPEA, in ACN (Method B) toafford compounds of formula iii, that can be subsequently coupled with asuitable α-haloacetate ester, such as iodoethylacetate orbromoethylacetate in a one-pot process using microwave irradiation toafford substituted thiophenes iv (Method C). Esters iv can be hydrolyzedusing a suitable base, such as NaOH in aqueous conditions usingcosolvents, such as THF and MeOH to afford carboxylic acids v (MethodD). Amides vi can be obtained by coupling of compounds v with ammoniausing a suitable coupling reagent, such as EDCI and HOBT in DCM (MethodE). Treatment of compounds vi with DMFDMA under microwave irradiationgives intermediate enamines, that are transformed to triazoles vii usinghydrazine in acetic acid under microwave irradiation.

Scheme 2 above shows a general route for preparing compounds of formula(iv). As shown in Scheme 2, viii is treated with ethyl thioacetate inthe presence of a suitable base, such as TEA in MeOH under elevatedtemperature to afford ix (Method G), which is subjected to Sandmeyerreaction using appropriate reagents, such as methylene iodide and amylnitrite in ACN (Method H). The product x is then oxidized to sulfone xiusing an appropriate oxidant, such as m-CPBA in DCM (Method I).Displacement of a sulfone is achieved using morpholine in THF atelevated temperature (Method J) to give xii. Suzuki coupling of thelatter compound with boronic acids is achieved using an appropriatecatalyst, such as Pd(PPh₃)₄, in the presence of a suitable base, such assodium carbonate in DME-water mixture under microwave conditions toafford compounds of formula iv (Method K). Transformation of esters ivto triazoles vii is carried out as described in Scheme 1.

Scheme 3 above shows a general route for preparing compounds of formula(xvii). As shown in Scheme 3, ester x is hydrolyzed using a suitablebase, such as sodium hydroxide in aqueous conditions, to give carboxylicacid xiii (Method D). Formation of amide xiv is done using anappropriate coupling reagent, such as EDCI and HOBT in DCM followed bytreatment with aqueous ammonia (Method E). Thioether xiv can be oxidizedto sulfone xv using a suitable oxidant, such as mCPBA in DCM (Method L).The latter compound is subjected to Suzuki coupling conditions with anappropriate combination of aryl boronic acid, Pd source, such asPd(dba)₂, ligand, such as dpePhos, and a base, such as potassiumphosphate in DME/DMA solvent mixture under microwave irradiation toafford advanced intermediate of formula xvi (Method M). Treatment ofsulfones xvi with neat substituted morpholines under elevatedtemperature affords amides of formula xvii (Method N). Transformation ofamides xvii to triazoles vii is carried out as described in Scheme 1.

Scheme 4 above shows a general route for preparing compounds of formula(xx). As shown in Scheme 4, acid v is treated with Boc protectedethylenediamine using standard coupling conditions, such as EDCI andHOBt in DCM. Protective group is removed using an appropriate acid, forexample TFA in DCM to give amide xviii (Method O). Cyclization of xviiiis achieved using suitable conditions, for example POCl₃ (Method P) andthe formed dihydroimidazole xix is oxidized to imidazole xx using asuitable oxidative method, such as Swern oxidation (Method Q).

Scheme 5 above shows an alternative route for preparing compounds offormula (xx). As shown in Scheme 5, acid v is transformed to an aldehydeusing a suitable synthetic sequence, for example through a coupling withN,O-dimethylhydroxylamine and subsequent reduction of the formed Weinrebamide with DIBAL (Method R). Aldehyde xxi is then treated with diamineand iodine (Method S) to form dihydroimidazole xix, which is oxidized toimidazole xx using a suitable oxidating method, such as Swern oxidation(Method Q).

Scheme 6 above shows an alternative route for preparing compounds offormula (xxv). As shown in Scheme 6, acid v is transformed to a ketoneusing a suitable synthetic sequence, for example through a coupling withN,O-dimethylhydroxylamine and subsequent treatment of the formed Weinrebamide with alkyllithium reagent (Method T). Ketone xxii is then treatedwith bromine in acetic acid under microwave irradiation (Method U) toform dibromoketone xxiii, which is reduced to bromoketone xxiv using asuitable method, such as diethyl phosphate and a base, such as TEA(Method V). Treatment of xxiv with formamide under microwave irradiationaffords the final imidazole xxv (Method W).

Scheme 7 above shows a general route for preparing compounds of formula(xxvi). As shown in Scheme 7, ketone xxii is treated with DMFDMA toafford an intermediate enamine followed by reaction with hydrazinehydrate in a suitable solvent, for example acetic acid to give pyrazolexxvi (Method X).

Scheme 8 above shows a general route for preparing compounds of formula(xxvii). As shown in Scheme 8, amide vi is treated with an azide source,for example sodium azide using a suitable Lewis acid, for examplesilicon tetrachloride in an appropriate solvent, such as acetonitrile togive tetrazole xxvii (Method Y).

Scheme 9 above shows a general route for preparing compounds of formula(xxxiv). As shown in Scheme 9, sulfone xi is treated withdimethoxybenzylamine using suitable conditions, for example TEA in THFat elevated temperature. Suzuki coupling of the latter compound withboronic acids is achieved using an appropriate catalyst, such asPd(PPh₃)₄, in the presence of a suitable base, such as sodium carbonatein DME-water mixture under microwave conditions to afford compounds offormula xxix (Method K). Deprotection of dimethoxybenzyl group isachieved with a suitable acid, such as TFA in DCM (Method AA). Aminesxxx are then subjected to Sandmeyer reaction using appropriate reagents,such as methylene iodide and amyl nitrite in ACN (Method H). Esters xxxican be hydrolyzed using a suitable base, such as NaOH in aqueousconditions using cosolvents, such as THF and MeOH to afford carboxylicacids (Method D) followed by coupling with ammonia using a suitablecoupling reagent, such as EDCI and HOBT in DCM to give amides xxxii(Method E). Treatment of amides xxxii with DMFDMA under microwaveirradiation gives intermediate examines that are transformed totriazoles xxxiii using hydrazine in acetic acid under microwaveirradiation (Method F). Compounds xxxiii can be then coupled withvinylstannanes under suitable conditions, for example Pd(PPh₃)₄, CuI,LiCl in dioxane under elevated temperature to give dihydropyrans xxxiv(Method AB). Hydrogenation of the latter compounds (Method AC) canafford tetrahydropyrans xxxv.

Scheme 10 above shows a general route for preparing compounds of formula(xl). As shown in Scheme 10, 2,4-dichlorothiazole-5-carbonitrile istransformed to xxxvii by treatment with morpholine (Method AD), which inturn is subjected to Suzuki coupling under microwave conditions usingstandard reagents, for example Pd(PPh₃)₂Cl₂, sodium carbonate inDME-ethanol-water mixture (Method K) to afford compounds of formulaxxxviii. Hydrolysis of the nitrile group is achieved using a strongacid, such as sulfuric acid (Method AE) to give amides xxxix, which aretransformed to triazoles xl using a two step procedure involvingtreatment with DMFDMA under microwave irradiation followed by hydrazinein acetic acid under microwave irradiation (Method F).

Compounds of formula (I), where X═O or X═N(R) can be prepared fromcompounds xliii. The synthesis of compounds of formula xliii is reportedin the literature (Fernandez, M.-C.; Castano, A.; Dominguez, E.;Escribano, A.; Jiang, D.; Jimenez, A.; Hong, E.; Homback, W. J.;Nisenbaum, E. S.; Rankl, N.; Tromiczak, E.; Vaught, G.; Zarrinmayeh, H.;Zimmerman, D. M. Bioorg. Med. Chem. Lett., 2006, 66, 5057-5061), and isoutlined in Scheme 11. Deprotonation of propionitriles xli followed bycondensation with carbon disulfide and subsequent quenching with methyliodide gives compounds of formula xlii. These compounds can be furtherconverted to furan ethyl carboxylates xliii by cyclocondensation withbromoethyl acetate. Similarly, treatment of xlii with N-alkyl glycineesters affords alkylated pyrrole ethyl carboxylates xliv. Compounds offormula xliii and xliv can be further elaborated as described in Schemes1-9 above.

5. Uses, Formulation and Administration

As discussed above, the present invention provides compounds that areuseful as inhibitors of PI3K enzymes, and thus the present compounds areuseful for treating proliferative, inflammatory, or cardiovasculardisorders such as tumor and/or cancerous cell growth mediated by PI3K.In particular, the compounds are useful in the treatment of cancers in asubject, including, but not limited to, lung and bronchus, prostate,breast, pancreas, colon and recum, thyroid, liver and intrahepatic bileduct, hepatocellular, gastric, glioma/glioblastoma, endometrial,melanoma, kidney, and renal pelvis, urinary bladder, utering corpus,uterine cervix, ovary, multiple myeloma, esophagus, acute myelogenousleukemia, chronic myelogenous leukemia, lymphocytic leukemia, myeloidleukemia, brain, oral cavity, and pharynx, small intestine, non-Hodgkinlymphoma, and villous colon adenoma.

In some embodiments, compounds of the invention are suitable for thetreatment of breast cancer, bladder cancer, colon cancer, glioma,glioblastoma, lung cancer, hepatocellular cancer, gastric cancer,melanoma, thyroid cancer, endometrial cancer, renal cancer, cervicalcancer, pancreatic cancer, esophageal cancer, prostate cancer, braincancer, or ovarian cancer.

In other embodiments, compounds of the invention are suitable for thetreatment of inflammatory and cardiovascular disorders including, butnot limited to, allergies/anaphylaxis, acute and chronic inflammation,rheumatoid arthritis; autoimmunity disorders, thrombosis, hypertension,cardiac hypertrophy, and heart failure.

Accordingly, in another aspect of the present invention, pharmaceuticalcompositions are provided, wherein these compositions comprise any ofthe compounds as described herein, and optionally comprise apharmaceutically acceptable carrier, adjuvant or vehicle. In certainembodiments, these compositions optionally further comprise one or moreadditional therapeutic agents.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative thereof. According to thepresent invention, a pharmaceutically acceptable derivative includes,but is not limited to, pharmaceutically acceptable prodrugs, salts,esters, salts of such esters, or any other adduct or derivative whichupon administration to a patient in need is capable of providing,directly or indirectly, a compound as otherwise described herein, or ametabolite or residue thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. A“pharmaceutically acceptable salt” means any non-toxic salt or salt ofan ester of a compound of this invention that, upon administration to arecipient, is capable of providing, either directly or indirectly, acompound of this invention or an inhibitorily active metabolite orresidue thereof. As used herein, the term “inhibitorily activemetabolite or residue thereof” means that a metabolite or residuethereof is also an inhibitor of PI3K.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersable products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, lower alkyl sulfonate and aryl sulfonate.

As described above, the pharmaceutically acceptable compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, adjuvant, or vehicle, which, as used herein, includes any andall solvents, diluents, or other liquid vehicle, dispersion orsuspension aids, surface active agents, isotonic agents, thickening oremulsifying agents, preservatives, solid binders, lubricants and thelike, as suited to the particular dosage form desired. Remington'sPharmaceutical Sciences, Sixteenth Edition, E. W. Martin (MackPublishing Co., Easton, Pa., 1980) discloses various carriers used informulating pharmaceutically acceptable compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, or potassiumsorbate, partial glyceride mixtures of saturated vegetable fatty acids,water, salts or electrolytes, such as protamine sulfate, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, woolfat, sugars such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; powderedtragacanth; malt; gelatin; talc; excipients such as cocoa butter andsuppository waxes; oils such as peanut oil, cottonseed oil; saffloweroil; sesame oil; olive oil; corn oil and soybean oil; glycols; such apropylene glycol or polyethylene glycol; esters such as ethyl oleate andethyl laurate; agar; buffering agents such as magnesium hydroxide andaluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline;Ringer's solution; ethyl alcohol, and phosphate buffer solutions, aswell as other non-toxic compatible lubricants such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releasingagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

In yet another aspect, a method for treating a proliferative,inflammatory, or cardiovascular disorder is provided comprisingadministering an effective amount of a compound, or a pharmaceuticalcomposition to a subject in need thereof. In certain embodiments of thepresent invention an “effective amount” of the compound orpharmaceutical composition is that amount effective for treating aproliferative, inflammatory, or cardiovascular disorder, or is thatamount effective for treating cancer. In other embodiments, an“effective amount” of a compound is an amount which inhibits binding ofPI3K and thereby blocks the resulting signaling cascades that lead tothe abnormal activity of growth factors, receptor tyrosine kinases,protein serine/threonine kinases, G protein coupled receptors andphospholipid kinases and phosphatases.

The compounds and compositions, according to the method of the presentinvention, may be administered using any amount and any route ofadministration effective for treating the disease. The exact amountrequired will vary from subject to subject, depending on the species,age, and general condition of the subject, the severity of theinfection, the particular agent, its mode of administration, and thelike. The compounds of the invention are preferably formulated in dosageunit form for ease of administration and uniformity of dosage. Theexpression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the compoundsand compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disease beingtreated and the severity of the disease; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention may be administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subjectbody weight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active, compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

While one or more of the inventive compounds may be used in anapplication of monotherapy to treat a disorder, disease or symptom, theyalso may be used in combination therapy, in which the use of aninventive compound or composition (therapeutic agent) is combined withthe use of one or more other therapeutic agents for treating the sameand/or other types of disorders, symptoms and diseases. Combinationtherapy includes administration of the therapeutic agents concurrentlyor sequentially. Alternatively, the therapeutic agents can be combinedinto one composition which is administered to the patient.

In one embodiment, the compounds of this invention are used incombination with other therapeutic agents, such as other inhibitors ofPI3K. In some embodiments, a compound of the invention is administeredin conjunction with a therapeutic agent selected from the groupconsisting of cytotoxic agents, radiotherapy, and immunotherapy. It isunderstood that other combinations may be undertaken while remainingwithin the scope of the invention.

Another aspect of the invention relates to inhibiting PI3K, activity ina biological sample or a patient, which method comprises administeringto the patient, or contacting said biological sample with a compound offormula I or a composition comprising said compound. The term“biological sample”, as used herein, generally includes in vivo, invitro, and ex vivo materials, and also includes, without limitation,cell cultures or extracts thereof; biopsied material obtained from amammal or extracts thereof; and blood, saliva, urine, feces, semen,tears, or other body fluids or extracts thereof.

Still another aspect of this invention is to provide a kit comprisingseparate containers in a single package, wherein the inventivepharmaceutical compounds, compositions and/or salts thereof are used incombination with pharmaceutically acceptable carriers to treatdisorders, symptoms and diseases where PI3K kinase plays a role.

EXPERIMENTAL PROCEDURES I. Preparation of Exemplary CompoundsDefinitions

-   AcOH acetic acid-   ACN acetonitrile-   ATP adenosine triphosphate-   BCA bicinchoninic acid-   BSA bovine serum albumin-   BOC tert-butoxycarbonyl-   m-CPBA m-chloroperbenzoic acid-   DCE dichloroethane-   DCM dichloromethane-   DDQ 2,3-dichloro-5,6-dicyano-1,4-benzoquinone-   DIPEA diisopropylethyl amine-   DMEM Dulbecco's Modified Eagle's Medium-   DMF N,N-dimethylformamide-   DMFDMA N,N-dimethylformamide dimethyl acetal-   DMSO dimethylsulfoxide-   DPPA diphenylphosphoryl azide-   DTT dithiothreitol-   dppf diphenylphosphinoferrocene-   EDCI N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride-   EDTA ethylenediaminetetraacetic acid-   EtOAc ethyl acetate-   EtOH ethanol-   FA formic acid-   FBS fetal bovine serum-   h hours-   HATU N,N,N′,N′-tetramethyl-o-(7-azabenzotriazole-1-yl)uronium    hexafluorophosphate-   HBTU o-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium    hexafluorophosphate-   HEPES N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)-   HOBT 1-hydroxybenzotriazole hydrate-   HRMS high resolution mass spectrum-   LAH lithium aluminum hydride-   LCMS liquid chromatography mass spectrum-   m/z mass to charge-   Me methyl-   MeOH methanol-   min minutes-   MS mass spectrum-   MTT methylthiazoletetrazolium-   MWI microwave irradiation-   PBS phosphate buffered saline-   PKA cAMP-dependent protein kinase-   rt room temperature-   TEA triethylamine-   TFFA trifluoroacetic anhydride-   THF tetrahydrofuran-   TMB 3,3′,5,5′-Tetramethylbenzidine-   WST    (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene    disulfonate sodium salt)

Analytical LC-MS Methods

LCMS Conditions

Spectra were run on a Phenominex Luna 5 μm C18 50×4.6 mm column on aHewlett-Packard HP1100 using the following gradients:

-   -   Method Formic Acid (FA): Acetonitrile containing 0 to 100        percent 0.1% formic acid in water (2.5 ml/min for a 3 minute        run).    -   Method Ammonium Acetate (AA): Acetonitrile containing 0 to 100        percent 10 mM ammonium acetate in water (2.5 ml/min for a 3        minute run).

Example 1 Synthesis of4-(2,4-dichlorophenyl)-2-morpholin-4-yl-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(Compound I-25)

Step 1: 3-morpholin-4-yl-3-thioxopropanenitrile

To a solution of 3-morpholin-4-yl-3-oxopropanenitrile (3.0 g, 19.5 mmol)in anhydrous THF (45 mL) was added Lawesson's reagent (4.2 g, 10.3mmol). The reaction mixture was allowed to stir at rt for 16 h and wasthen concentrated to small volume. A solid precipitated and wasfiltered. The solid was washed with diethyl ether to give3-morpholin-4-yl-3-thioxopropanenitrile (2.3 g, 70%). LCMS: (AA) ES+171.2. ¹H NMR (400 MHz, d₁-chloroform) δ: 4.29-4.27 (m, 2H), 4.00 (s,2H) and 3.83-3.81 (m, 6H).

Step 2 and 3: Ethyl4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxylate

A solution of 3-morpholin-4-yl-3-thioxopropanenitrile (0.150 g, 0.88mmol) in anhydrous ACN (1.2 mL) was cooled to 0° C. To this cooledsolution was added of 2,4-dichlorobenzoyl chloride (0.149 mL, 1.06 mmol)and DIPEA (0.161 mL, 0.93 mmol). The mixture was allowed to stir at itunder nitrogen for 15 min before ethyl iodoacetate (0.110 mL, 0.93 mmol)and DIPEA (0.161 mL, 0.93 mmol) were added. The reaction mixture wassubjected to MWI at 140° C. for 10 min. The mixture was allowed to cooland was concentrated. The residue was purified by column chromatographyto give ethyl4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxylate(0.1 g, 28%). LCMS: (AA) ES+ 411. ¹H NMR (400 MHz, d₁-chloroform) δ:7.49 (d, 1H), 7.33 (dd, 1H), 7.20 (d, 1H), 4.16-4.08 (m, 2H), 3.89-3.85(m, 4H), 3.66-3.63 (m, 4H) and 1.12 (t, 3H).

Step 4:4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxylicacid

To a solution of ethyl4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxylate(0.060 g, 0.15 mmol) in THF/MeOH/water (2:1:2) (5 mL) was added sodiumhydroxide (0.061 g, 1.5 mmol). The reaction mixture was allowed to stirat rt for 20 h and was concentrated. The residue was acidified with 1NHCl and was extracted with EtOAc. The organic solutions were combined,washed with brine, dried over MgSO₄, filtered and concentrated to give4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxylicacid (0.034 g, 62%). LCMS: (FA) ES+ 382.9. ¹H NMR (400 MHz, d₆-DMSO) δ:7.75 (d, 1H), 7.50 (dd, 1H), 7.39 (d, 1H), 3.79-3.74 (m, 4H) and3.61-3.55 (m, 4H).

Step 5:4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxamide

4-Cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxylicacid (0.050 g, 0.13 mmol), HOBT (0.037 g, 0.274 mmol) and EDCI (50 mg,0.261 mmol) were suspended in DCM (6.5 mL). After 30 min the reagentsdissolved. To the resulting solution was added concentrated aqueousammonia (0.26 mL, 6.5 mmol) and the solution was allowed to stirvigorously at rt overnight. The reaction mixture was concentrated andthe residue was diluted with 1N HCl and extracted with EtOAc. Theorganic solutions were combined, washed with sat NaHCO₃ and brine, driedover MgSO₄ filtered and concentrated to give a brown solid. The solidwas triturated with hexanes and cold EtOAc to give4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxamide(0.024 g, 49%). LCMS: (AA) ES+ 382, ES− 380. ¹H NMR (400 MHz, d₆-DMSO)δ: 7.78 (s, 1H), 7.54 (d, 2H), 7.43 (d, 2H), 3.79-3.77 (m, 4H) and3.55-3.52 (m, 4H).

Step 6: Synthesis of4-(2,4-dichlorophenyl)-2-morpholin-4-yl-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile

A mixture of4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxamide(31.6 mg, 0.0000827 mol) in 1,1-dimethoxy-N,N-dimethylmethanamine (1.0mL, 0.0075 mol) was flushed with argon and then irradiated in microwaveat 160° C. for 60 min. The reaction mixture was concentrated to give4-cyano-3-(2,4-dichlorophenyl)-N-((dimethylamino)methylene)-5-morpholinothiophene-2-carboxamideas intermediate (31 mg, 86%). LCMS: (FA) ES+ 437.13. ¹H NMR (400 MHz,d₆-DMSO) δ: 8.34 (s, 1H), 7.69 (d, 1H), 7.47 (d, 1H), 7.33 (d, 1H),3.79-3.74 (m, 4H), 3.59-3.54 (m, 4H), 3.07 (s, 3H), 2.72 (s, 3H). To theabove intermediate in AcOH (1.8 mL, 0.032 mol) was added hydrazinehydrate (50 mg, 0.001 mol). The mixture was flushed with argon and thenirradiated in microwave at 120° C. for 20 min. The mixture wasconcentrated to remove most of the solvent. Water was added to theresidue and the precipitate was collected, washed with water, dried toafford4-(2,4-dichlorophenyl)-2-morpholino-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(12.8 mg, 55%). LCMS: (FA) ES+ 406.11, ES− 404.21, ¹H NMR (400 MHz,d₆-DMSO) δ: 14.00 (s, 1H), 8.43 (s, 1H), 7.74 (d, 1H), 7.49 (d, 1H),7.41 (d, 1H), 3.82-3.76 (m, 4H), 3.56-3.51 (m, 4H).

Compounds in the following table were prepared from the appropriatestarting materials in a method analogous to that of Example 1:

4 LCMS: (FA) ES+ 449.95. 8 LCMS: (FA) ES+ 387.99. 10 LCMS: (FA) ES+ 402.12 LCMS: (FA) ES+ 434. 18 LCMS: (FA) ES+ 440.14. 26 LCMS: (FA) ES+ 434.

Example 24-(2,4-dichlorophenyl)-2-(2-(fluoromethyl)morpholino)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(Compound I-11)

Step 1: Ethyl 3-amino-4-cyano-5-(methylsulfanyl)thiophene-2-carboxylate

A mixture of [bis(methylsulfanyl)methylene]malononitrile (40 g, 230mmol), ethylthioglycolate (29 g, 230 mmol) and TEA (24 mL, 173 mmol) inMeOH (600 mL) was allowed to stir at reflux for 2 h. The reactionmixture was allowed to cool overnight and the precipitate was filteredoff, washed with cold MeOH (3×50 mL) to give ethyl3-amino-4-cyano-5-(methylsulfanyl)thiophene-2-carboxylate (52.4 g, 99%).LCMS: (FA) ES+ 275.

Step 2: Ethyl 4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxylate

Ethyl 3-amino-4-cyano-5-(methylsulfanyl)thiophene-2-carboxylate (10 g,41.3 mmol) was dissolved in acetonitrile (50 mL) under atmosphere ofargon. Diiodomethane (11.6 mL, 0.144 mol) was added and the mixture washeated at 40° C. Isoamyl nitrite (12.1 g, 0.103 mol) was added and thereaction was allowed to cool to room temperature and stirred for 2hours. Mixture was cooled down at 0° C., diluted with hexane (50 mL) andthe precipitate was filtered off, washed with 10:1 hexane-acetonitrilemixture (10 mL), 3:1 hexane-ether (10 mL) and hexane (10 mL). Theprecipitate was dried to afford ethyl4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxylate (6.90 g, 45%).LCMS: (FA) ES+ 354. ¹H NMR (400 MHz, d₁-chloroform) δ: 4.38 (q, 2H),2.70 (s, 3H), 1.40 (t, 3H).

Step 3: Ethyl 4-cyano-3-iodo-5-(methylsulfonyl)thiophene-2-carboxylate

Ethyl 4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxylate (7.2 g,20.4 mmol) was dissolved in DCM (200 mL) and THF (100 mL) and m-CPBA(9.14 g, 40.8 mmol) was added. The reaction mixture was stirred at rtovernight. Sodium sulfite (5.14 g, 40.8 mmol) was added, stirred for 10minutes followed by addition of potassium carbonate (8.45, 61.2 mmol).The suspension was stirred at rt for 1 hour and filtered through celite,washed with DCM and the solvent was evaporated to afford ethyl3-iodo-4-cyano-5-(methylsulfonyl)thiophene-2-carboxylate (6.80 g, 78%).LCMS: (FA) ES+ 386. ¹H NMR (400 MHz, d₁-chloroform) δ: 4.45 (q, 2H),3.38 (s, 3H), 1.43 (t, 3H).

Step 4: Ethyl4-cyano-5-(2-(fluoromethyl)morpholino)-3-iodothiophene-2-carboxylate

A mixture of ethyl4-cyano-3-iodo-5-(methylsulfonyl)thiophene-2-carboxylate (718 mg,0.00186 mol), 2-(fluoromethyl)morpholine hydrochloride (348 mg, 0.00224mol) and TEA (1.0 mL, 0.0072 mol) in THF (7.1 mL, 0.088 mol) was flushedwith argon and then heated at 90° C. overnight. The reaction mixture wasconcentrated and purified by column chromatography to give ethyl4-cyano-5-(2-(fluoromethyl)morpholino)-3-iodothiophene-2-carboxylate(288 mg, 36.4%). LCMS: (FA) ES+ 425, ¹H NMR (400 MHz, d₁-chloroform) δ:4.57 (d, 1H), 4.46 (d, 1H), 4.34 (q, 2H), 4.12-4.05 (m, 1H), 4.02-3.77(m, 4H), 3.34-3.25 (m, 1H), 3.22-3.14 (m, 1H), 1.37 (t, 3H).

Step 5: ethyl4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxylate

A mixture of ethyl4-cyano-5-(2-(fluoromethyl)morpholino)-3-iodothiophene-2-carboxylate(288 mg, 0.000679 mol), 2,4-dichlorophenylboronic acid (262 mg, 0.00137mol), tetrakis(triphenylphosphine)palladium(0) (51 mg, 0.000044 mol) andsodium carbonate (220 mg, 0.0021 mol) in water (3 mL, 0.1 mol) and1,2-dimethoxyethane (10 mL, 0.1 mol) was flushed with argon and thenirradiated in microwave at 140° C. for 12 min. The reaction mixture wasfiltered and concentrated in vacuum to afford ethyl4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxylate(300 mg, 99%). LCMS: (FA) ES+ 443.15.

Step 6:4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxylicacid

A mixture of ethyl4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxylate(300 mg, 0.00054 mol), Sodium hydroxide (0.12 g, 0.0030 mol) in EtOH(10.0 mL, 0.171 mol) and water (3.0 mL, 0.17 mol) was stirred at rtuntil completion by LCMS. The mixture was concentrated and the residuewas acidified by 1NHCl. The precipitate was collected and dried in airand vacuum to afford4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxylicacid (225 mg, 99%). LCMS: (FA) ES+ 415, ES− 413.

Step 7:4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxamide

A mixture of4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxylicacid (68.0 mg, 0.000164 mol), 33% ammonium hydroxide (33:67,ammonia:water, 0.1416 mL), EDCI (94.2 mg, 0.000491 mol) and 1-HOBT (66.4mg, 0.000491 mol) in DCM (7 mL, 0.1 mol) was stirred at rt overnight.The reaction mixture was diluted with DCM and washed with water, theorganic layer was dried and purified by column chloromatography toafford4-cyano-3-(2,4-dichlorophenyl)-5-(2-(fluoromethyl)morpholino)thiophene-2-carboxamide(38 mg, 56%). LCMS: (FA) ES+ 414, ES− 412; ¹H NMR (400 MHz,d₁-chloroform) δ: 7.59 (d, 1H), 7.42 (dd, 1H), 7.31 (d, 1H), 6.00 (s,1H), 4.50 (dd, 2H), 4.12-4.08 (m, 2H), 4.00-3.78 (m, 4H), 3.36-3.25 (m,1H), 3.22-3.15 (m, 1H)

Step 8:4-(2,4-dichlorophenyl)-2-(2-(fluoromethyl)morpholino)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile

A solution of4-cyano-3-(2,4-dichlorophenyl)-5-[2-(fluoromethyl)morpholin-4-yl]thiophene-2-carboxamide(38 mg, 0.000092 mol) in 1,1-dimethoxy-N,N-dimethylmethanamine (2.0 mL,0.015 mol) was irradiated in microwave at 160° C. for 40 min. Thesolvent was removed and the residue was dissolved in AcOH (1.6 mL, 0.028mol). Hydrazine monohydrate (0.3 mL, 0.006 mol) was added and themixture was irradiated in microwave at 120° C. for 10 min. The reactionmixture was concentrated and the residue was treated with water, thesolid was collected, dried to afford4-(2,4-dichlorophenyl)-2-(2-(fluoromethyl)morpholino)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(23.5 mg, 60%). LCMS: (FA) ES+ 438, ES− 436; ¹H NMR (400 MHz, d₆-DMSO)δ: 8.01 (s, 1H), 7.57 (d, 1H), 7.39 (dd, 1H), 7.32 d, 1H), 4.59 (t, 1H),4.47 (t, 1H), 4.08-4.01 (m, 1H), 4.00-3.81 (m, 5H), 3.32-3.20 (m, 1H),3.18-3.09 (m, 1H).

Compounds in the following table were prepared from the appropriatestarting materials in a method analogous to that of Example 2:

3 LCMS: (FA) ES+ 434. 7 LCMS: (FA) ES+ 438. 9 LCMS: (FA) ES+ 434. 14LCMS: (FA) ES+ 434. 15 LCMS: (FA) ES+ 420. 17 LCMS: (FA) ES+ 368. 32LCMS: (FA) ES+ 438. 33 LCMS: (FA) ES+ 388. 35 LCMS: (FA) ES+ 434.

Example 3 Synthesis of4-(2,4-dichlorophenyl)-2-[2-(hydroxymethyl)morpholin-4-yl]-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(Compound I-27)

Step 1: 4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxylic acid

To a solution of ethyl4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxylate (3.40 g, 10mmol) in tetrahydrofuran (80 mL) and water (16 mL) was added a solutionof 1.00M sodium hydroxide in water (30 mL). The solution was allowed tostir overnight. The reaction was quenched with a solution of 1N hydrogenchloride in water (50 mL) and diluted with water (400 mL). The resultantprecipitate was filtered, washed with water (2×100 mL) and dried in avacuum oven to give4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxylic acid (2.6 g,79%) as a white solid. LCMS: (FA) ES+ 326. ¹H NMR (400 MHz, d₆-DMSO) δ:14.1-13.8 (bs, 1H), 2.75 (s, 3H).

Step 2: 4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxamide

To a suspension of4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxylic acid (2.85 g,7.93 mmol) in methylene chloride (30 mL), were addedN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (3.28 g,17.1 mmol) and 1-hydroxybenzotriazole (2.27 g, 16.8 mmol). The reactionmixture was stirred at room temperature for two hours and ammoniumhydroxide (15.4 mL) was added and the biphasic mixture was stirred atroom temperature for two hours. Water (100 mL), methanol (50 mL),methylene chloride (200 mL) was added. The organic layer was removed.The aqueous layer was extracted five times with a solution of 20%methanol in methylene chloride (100 mL). The combined organic extractswere dried over anhydrous magnesium sulfate, filtered and concentratedto the title compound as dark red oil (1.47 g, 57%). LCMS: (FA) ES+ 325.

Step 3: 4-cyano-3-iodo-5-(methylsulfonyl)thiophene-2-carboxamide

To a solution of4-cyano-3-iodo-5-(methylsulfanyl)thiophene-2-carboxamide (1.46 g, 4.50mmol) in methylene chloride (60 mL), tetrahydrofuran (20 mL),N,N-dimethylformamide (20 mL), m-chloroperbenzoic acid (5.05 g, 22.5mmol) was added and the mixture was stirred at room temperatureovernight. The methylene chloride was removed in vacuo. The remainingresidue was diluted with ethyl acetate (200 mL) and washed three timeswith a solution of 1.00M sodium hydroxide in water (50 mL). The organicphase was removed and the aqueous phase was extracted five times withethyl acetate (100 mL). The combined organic extracts were washed twicewith a solution of 1.00M sodium hydroxide in water (50 mL). The organicextracts were concentrated in vacuo. The residue was suspended in water(100 mL) and a precipitate formed. The precipitate was filtered, washedwith water (40 mL), hexanes (100 mL), and dried in a vacuum oven to givethe title compound as a white solid (1.00 g, 62%). LCMS: (FA) ES+ 357.¹H NMR (400 MHz, d₆-DMSO) δ: 8.16 (s, 2H), 3.56 (s, 3H).

Step 4:4-cyano-3-(2,4-dichlorophenyl)-5-(methylsulfonyl)thiophene-2-carboxamide

4-cyano-3-iodo-5-(methylsulfonyl)thiophene-2-carboxamide (0.500 g, 1.40mmol), bis(dibenzylideneacetone)palladium (0.040 g, 0.0700 mmol),bis(2-diphenylphosphinophenyl)ether (0.057 g, 0.100 mmol), and potassiumphosphate (0.596 g, 2.81 mmol) were suspended in 1,2-dimethoxyethane(10.0 mL) and N,N-dimethylacetamide (5 mL). The suspension was flushedwith argon and the reaction mixture was irradiated in microwave at 150°C. (300 watts) for three hours. The reaction mixture was concentrated invacuo and column chromatography was performed to yield the titlecompound (0.080 g, 14%) as beige foam. ¹H NMR (400 MHz, d₄-methanol) δ:7.71-7.69 (m, 1H), 7.52-7.50 (m, 2H), 3.46 (s, 3H).

Step 5:4-cyano-3-(2,4-dichlorophenyl)-5-[2-(hydroxymethyl)morpholin-4-yl]thiophene-2-carboxamide

4-cyano-3-(2,4-dichlorophenyl)-5-(methylsulfonyl)thiophene-2-carboxamide(0.026 g, 0.069 mmol) was dissolved in 2-hydroxymethylmorpholine (0.300g, 2.56 mmol) and the solution was heated at 60° C. overnight. Theresidue was concentrated in vacuo and column chromatography wasperformed and yielded4-cyano-3-(2,4-dichlorophenyl)-5-[2-(hydroxymethyl)morpholin-4-yl]thiophene-2-carboxamide(0.020 g, 66%) as a white solid. LCMS: (FA) ES+ 412. ¹H NMR (400 MHz,d₄-methanol) δ: 7.76 (s, 1H), 7.49 (d, 1H), 7.40 (d, 1H), 4.08-3.92 (m,3H), 3.84-3.72 (m, 2H), 3.69-3.59 (m, 2H), 3.15-3.08 (m, 2H).

Step 6: Synthesis of4-(2,4-dichlorophenyl)-2-[2-(hydroxymethyl)morpholin-4-yl]-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile

A mixture of4-cyano-3-(2,4-dichlorophenyl)-5-[2-(hydroxymethyl)morpholin-4-yl]thiophene-2-carboxamide(115 mg, 0.279 mmol) in 1,1-dimethoxy-N,N-dimethylmethanamine (3.44 mL,0.025 mol) was flushed with argon and then irradiated in microwave at160° C. for 60 min. The reaction mixture was concentrated to give4-cyano-3-(2,4-dichlorophenyl)-N-[(1E)-(dimethylamino)methylene]-5-[2-(hydroxymethyl)morpholin-4-yl]thiophene-2-carboxamide(130 mg, 99%). LCMS: (FA) ES+ 467. To the above intermediate in AcOH(4.3 mL, 0.076 mol) was added hydrazine hydrate (100 mg, 0.003 mol). Themixture was flushed with argon and then irradiated in microwave at 120°C. for 20 min. The mixture was concentrated to remove the solvent. Theresidue was purified using ISCO chromatography on silica gel, elution1:1 hexane-ethyl acetate to ethylacetate to afford4-(2,4-dichlorophenyl)-2-[2-(hydroxymethyl)morpholin-4-yl]-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(90 mg, 70%). LCMS: (FA) ES+ 436, ¹H NMR (400 MHz, d₄-methanol) δ: 8.28(s, 1H), 7.55 (d, 1H), 7.41 (dd, 1H), 7.34 (d, 1H), 4.12-3.72 (m, 5H),3.68-3.61 (m, 2H), 3.30-3.20 (m, 1H), 3.12-3.02 (m, 1H).

Compounds in the following table were prepared from the appropriatestarting materials in a method analogous to that of Example 3:

20 LCMS: (FA) ES+ 436. 24 LCMS: (FA) ES+ 492. 28 LCMS: (FA) ES+ 436. 31LCMS: (FA) ES+ 478. 36 LCMS: (FA) ES+ 450.

Example 4 Synthesis of4-(2,4-dichlorophenyl)-5-(1H-imidazol-2-yl)-2-morpholine-3-carbonitrile(Compound I-22)

Step 1:N-(2-aminoethyl)-4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxamide

To a solution of4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxylicacid (3.10 g, 8.2 mmol) and N-(2-aminoethyl)(tert-butoxy)carboxamide(1.98 gr, 12.3 mmol) in methylene chloride (60 mL) was addedN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (3.15 gr,16.4 mmol) and 1-hydroxybenzotriazole (2.22 gr, 16.4 mmol) at roomtemperature. The solution was allowed to stir overnight. The reactionwas diluted with methylene chloride and water. The layers were separatedand the organic layer was dried over anhydrous sodium sulfate, filteredand concentrated in vacuo. The residue was purified by columnchromatography to give the intermediate boc-protected product as ayellow solid. The material was dissolved in dioxane (4 mL) and asolution 4N HCl in dioxane (9 mL) was added. The solution was stirred atroom temperature for 30 minutes. The solvents were evaporated and theresidue was dried under vacuum overnight. The product was converted tothe free base by suspension in methylene chloride followed by additionof sodium carbonate solution. The layers were separated and the aqueouslayer was extracted twice with methylene chloride. The combined organicextracts were dried over anhydrous sodium sulfate, filtered andconcentrated in vacuo to afford the product as a free base (2.55 gr,67%). LCMS: (FA) ES+ 463. ¹H NMR (400 MHz, d₁-chloroform) δ: 7.59-7.55(m, 1H), 7.43-7.30 (m, 2H), 3.90-3.82 (m, 4H), 3.64-3.57 (m, 4H),3.53-2.85 (m, 4H).

Step 2:4-(2,4-dichlorophenyl)-5-(4,5-dihydro-1H-imidazol-2-yl)-2-morpholin-4-ylthiophene-3-carbonitrile

To a suspension ofN-(2-aminoethyl)-4-cyano-3-(2,4-dichlorophenyl)-5-morpholin-4-ylthiophene-2-carboxamide(2.50 g, 5.90 mmol) in toluene (20 mL) was added phosphoryl chloride(4.70 mL, 50.0 mmol) and the mixture was stirred under microwaveirradiation at 120° C. for 25 minutes. The reaction mixture wasevaporated and the residue was suspended in methylene chloride and thenquenched with ice water. The aqueous layer was basified by addition ofKOH, the layers were separated and the aqueous layer was extracted againwith methylene chloride. The organic layers were combined and washedwith brine, dried over anhydrous sodium sulfate, filtered andconcentrated in vacuo to afford the crude product (2.3 gr, 77%). LCMS:(FA) ES+ 408. ¹H NMR (400 MHz, d₆-DMSO) δ: 7.78-7.75 (m, 1H), 7.52-7.49(m, 2H), 3.79-3.73 (m, 4H), 3.63-3.55 (m, 2H), 3.54-3.48 (m, 4H),3.20-3.11 (m, 2H).

Step 3:4-(2,4-dichlorophenyl)-5-(1H-imidazol-2-yl)-2-morpholine-3-carbonitrile

To a solution of dimethyl sulfoxide (2.50 mL, 35.3 mmol) in methylenechloride (100 mL) at −78° C. under an atmosphere of argon was addedoxalyl chloride (2.98 mL, 35.3 mmol). The solution was stirred for 40minutes, then a solution of4-(2,4-dichlorophenyl)-5-(4,5-dihydro-1H-imidazol-2-yl)-2-morpholin-4-ylthiophene-3-carbonitrile(1.45 g, 3.03 mmol) in methylene chloride (40 mL) was added dropwise andthe reaction was stirred for 1 hour at −78° C. Triethylamine (14.0 mL,100 mmol) was added and stirring was continued at −78° C. for 1 hour.Then added 33% ammonium hydroxide (10 mL), and the mixture was allowedto warm to room temperature. Added sodium bicarbonate solution, thelayers were separated and the organic extract was dried over anhydroussodium sulfate, filtered and concentrated in vacuo to afford the crudeproduct. The residue was purified by column chromatography to give theproduct as a yellow solid (0.90 gr, 75%) which was dried overnight at39° C. in a vacuum oven LCMS: (FA) ES+ 406. ¹H NMR (400 MHz,d₁-chloroform) δ: 8.20-8.11 (s, 1H), 7.65-7.62 (m, 1H), 7.45-7.41 (m,1H), 7.37-7.33 (m, 1H), 7.03-7.00 (m, 1H), 6.85-6.81 (m, 1H), 3.90-3.84(m, 4H), 3.62-3.55 (m, 4H).

Compounds in the following table were prepared from the appropriatestarting materials in a method analogous to that of Example 4:

16 LCMS: (FA) ES+ 401. 30 LCMS: (FA) ES+ 437.

Example 5 Synthesis of4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)-2-morpholinothiophene-3-carbonitrile(Compound I-13)

Step 1:5-acetyl-4-(2,4-dichlorophenyl)-2-morpholinothiophene-3-carbonitrile

A mixture of4-cyano-3-(2,4-dichlorophenyl)-5-morpholinothiophene-2-carboxylic acid(508.0 mg, 0.001326 mol) in thionyl chloride (1.7 mL, 0.024 mol) andtoluene (10 mL, 0.098 mol) was heated at 80° C. for 1 h. The mixture wasevaporated in vacuum and the residue was coevaporated with toluene for 3times to afford acid chloride as dark brown oil. This oil was dissolvedin DCM (3 mL, 0.05 mol) and added to TEA (1.3 mL, 0.0093 mol) andN,O-dimethylhydroxylamine hydrochloride (282 mg, 0.00289 mol) in DCM (9mL, 0.1 mol) at 0° C. The mixture was stirred for 1 h at the sametemperature. The reaction mixture was washed with water and dried oversodium sulfate. The organic layer was evaporated to afford4-cyano-3-(2,4-dichlorophenyl)-N-methoxy-N-methyl-5-morpholinothiophene-2-carboxamide.To the above intermediate (207.0 mg, 0.0004856 mol) in THF (21 mL, 0.26mol) was added diisobutylaluminum hydride in hexane solution (1M, 0.1ml, 0.0019 mol) at −78° C. and the mixture was stirred at the sametemperature for 30 min. The reaction mixture was raised to 0° C. for 5min and cooled to −78° C. again. Quenched by MeOH and water and thenpurified by column chromatography to afford4-(2,4-dichlorophenyl)-5-formyl-2-morpholinothiophene-3-carbonitrile(115 mg, 65%). LCMS: (FA) ES+ 367. ¹H NMR (400 MHz, d₁-chloroform) δ:9.23 (s, 1H), 7.55 (d, 1H), 7.37 (dd, 1H), 7.30 (d, 1H), 3.88-3.84 (m,4H), 3.74-3.70 (m, 4H), 1.90 (s, 3H).

Step 2:4-(2,4-dichlorophenyl)-5-(4-methyl-4,5-dihydro-1H-imidazol-2-yl)-2-morpholinothiophene-3-carbonitrile

To4-(2,4-dichlorophenyl)-5-formyl-2-morpholin-4-ylthiophene-3-carbonitrile(52.0 mg, 0.000142 mol) in tert-butyl alcohol (0.75 g, 0.010 mol) wasadded 1,2-diaminopropane (12.9 mg, 0.000174 mol) and the mixture wasstirred for 30 min. Iodine (51 mg, 0.00020 mol) and potassium carbonate(65.7 mg, 0.000475 mol) was added the mixture was stirred at 70° C. for3 hr. The reaction mixture was quenched by saturated sodium bicarbonatesolution and brine. The organic layer was collected and purified bycolumn chromatography to afford4-(2,4-dichlorophenyl)-5-(4-methyl-4,5-dihydro-1H-imidazol-2-yl)-2-morpholinothiophene-3-carbonitrile(19 mg, 32%). LCMS: (FA) ES+ 421. ¹H NMR (400 MHz, d₁-chloroform) δ:7.60 (s, 1H), 7.50-7.40 (m, 1H), 7.40-7.30 (m, 1H), 4.20-4.05 (m, 1H),3.90-3.75 (m, 5H), 3.60-3.70 (m, 4H), 3.38-3.20 (m, 1H), 1.25 (s, 3H).

Step 3:4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)-2-morpholinothiophene-3-carbonitrile

To DMSO (32.0 uL, 0.000451 mol) in DCM (3 mL, 0.05 mol) at −78° C. wasadded oxalyl chloride (38.2 uL, 0.000451 mol) and the mixture wasstirred for 30 min.4-(2,4-dichlorophenyl)-5-(4-methyl-4,5-dihydro-1H-imidazol-2-yl)-2-morpholinothiophene-3-carbonitrile(19 mg, 0.000045 mol) in DCM (1.5 mL, 0.023 mol) was added. After 1 h,TEA (0.188 mL, 0.00135 mol) was added and the mixture was stirred foranother 30 min. 33% ammonium hydroxide (33:67, ammonia:Water, 0.117 mL)was added and the reaction mixture was warmed to room temperature. Themixture was purified by column chromatography to afford4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)-2-morpholinothiophene-3-carbonitrile(4.04 mg, 21%). LCMS: (FA) ES+ 419.13, ES− 417.17. ¹H NMR (400 MHz,d₁-chloroform) δ: 7.61 (d, 1H), 7.42 (dd, 1H), 7.34 (d, 1H), 6.61 (s,1H), 3.88-3.84 (m, 4H), 3.62-3.57 (m, 4H), 2.21 (s, 3H).

Compounds in the following table were prepared from the appropriatestarting materials in a method analogous to that of Example 5:

2 LCMS: (FA) ES+ 473.

Example 6 Synthesis of4-(2,4-dichlorophenyl)-5-(1H-imidazol-5-yl)-2-morpholinothiophene-3-carbonitrile(Compound I-19)

Step 1:5-acetyl-4-(2,4-dichlorophenyl)-2-morpholinothiophene-3-carbonitrile

A mixture of4-cyano-3-(2,4-dichlorophenyl)-5-morpholinothiophene-2-carboxylic acid(508.0 mg, 0.001326 mol) in thionyl chloride (1.7 mL, 0.024 mol) andtoluene (10 mL, 0.098 mol) was heated at 80° C. for 1 h. The mixture wasevaporated in vacuum and the residue was coevaporated with toluene for 3times to afford acid chloride as dark brown oil. This oil was dissolvedin DCM (3 mL, 0.05 mol) and added to TEA (1.3 mL, 0.0093 mol) andN,O-dimethylhydroxylamine hydrochloride (282 mg, 0.00289 mol) in DCM (9mL, 0.1 mol) at 0° C. The mixture was stirred for 1 h at the sametemperature. The reaction mixture was washed with water and dried oversodium sulfate. The organic layer was evaporated to afford4-cyano-3-(2,4-dichlorophenyl)-N-methoxy-N-methyl-5-morpholinothiophene-2-carboxamide.To the above intermediate in THF (50 mL, 0.6 mol) was addedmethyllithium in diethyl ether solution (1.4M) (1.7 ml, 0.00244 mol) at−78° C. After about 15 min, the reaction mixture was quenched byammonium chloride solution and extracted with ethyl acetate. The organiclayer was washed with brine and purified by column chromatography toafford5-acetyl-4-(2,4-dichlorophenyl)-2-morpholinothiophene-3-carbonitrile(397 mg, 78%). LCMS: (FA) ES+ 381.05. ¹H NMR (400 MHz, d₁-chloroform) δ:7.60 (d, 1H), 7.40 (dd, 1H), 7.28 (dd, 1H), 3.90-3.83 (m, 4H), 3.71-3.64(m, 4H), 1.90 (s, 3H).

Step 2:5-(2,2-dibromoacetyl)-4-(2,4-dichlorophenyl)-2-morpholinothiophene-3-carbonitrile

A solution of5-acetyl-4-(2,4-dichlorophenyl)-2-morpholin-4-ylthiophene-3-carbonitrile(50.0 mg, 0.000131 mol) and bromine (42 mg, 0.00026 mol) in AcOH (1.0mL, 0.018 mol) was flushed with argon and then irradiated in microwaveat 120° C. for 5 min. The reaction mixture was concentrated and theresidue was suspended in water. The solid was collected to gave5-(2,2-dibromoacetyl)-4-(2,4-dichlorophenyl)-2-morpholinothiophene-3-carbonitrile(46.9 mg, 66%). LCMS: (FA) ES+ 538.87; ¹H NMR (400 MHz, d₁-chloroform)δ: 7.82 (d, 1H), 7.45 (dd, 1H), 7.33 (d, 1H), 5.78 (s, 1H), 4.00-3.85(m, 4H), 3.80-3.70 (m, 4H).

Step 3:5-(2-bromoacetyl)-4-(2,4-dichlorophenyl)-2-morpholinothiophene-3-carbonitrile

To a mixture of5-(dibromoacetyl)-4-(2,4-dichlorophenyl)-2-morpholin-4-ylthiophene-3-carbonitrile(46.9 mg, 0.0000870 mol) in THF (0.78 mL, 0.0096 mol) was added diethylphosphite (13.7 uL, 0.000107 mol) and TEA (14.9 uL, 0.000107 mol) at 0°C. After addition, the mixture was warmed to rt after 20 min. Thereaction mixture was evaporated and the residue was suspended inice/water. The solid was filtered and purified by column chromatographyto afford5-(2-bromoacetyl)-4-(2,4-dichlorophenyl)-2-morpholinothiophene-3-carbonitrile(12 mg, 30%). LCMS: (FA) ES+ 460.88; ¹H NMR (400 MHz, d₁-chloroform) δ:7.59 (d, 1H), 7.42 (dd, 1H), 7.33 (d, 1H), 3.90-3.85 (m, 4H), 3.75-3.69(m, 5H), 3.62 (d, 1H).

Step 4:4-(2,4-dichlorophenyl)-5-(1H-imidazol-5-yl)-2-morpholinothiophene-3-carbonitrile

5-(Bromoacetyl)-4-(2,4-dichlorophenyl)-2-morpholin-4-ylthiophene-3-carbonitrile(50 mg, 0.0001 mol) in formamide (1.3 mL, 0.033 mol) was flushed withargon and then irradiated in microwave at 180° C. for 30 min. Thereaction mixture was concentrated and the residue was purified bypreparative HPLC to afford4-(2,4-dichlorophenyl)-5-(1H-imidazol-5-yl)-2-morpholinothiophene-3-carbonitrile(2.8 mg, 6%). LCMS: (FA) ES+ 405.18, ES− 403.27; ¹H NMR (400 MHz,d₄-methanol) δ: 7.66 (d, 1H), 7.60 (s, 1H), 7.46 (dd, 1H), 7.35 (d, 1H),6.27 (s, 1H), 3.89-3.40 (m, 4H), 3.55-3.49 (m, 4H).

Example 7 Synthesis of4-(2,4-dichlorophenyl)-2-morpholino-5-(1H-pyrazol-5-yl)thiophene-3-carbonitrile(Compound I-21)

Step 1:4-(2,4-dichlorophenyl)-2-morpholino-5-(1H-pyrazol-5-yl)thiophene-3-carbonitrile

A solution of5-acetyl-4-(2,4-dichlorophenyl)-2-morpholin-4-ylthiophene-3-carbonitrile(52.3 mg, 0.000137 mol) in 1,1-dimethoxy-N,N-dimethylmethanamine (1.0mL, 0.0075 mol) was flushed with argon and irradiated in microwave at160° C. for 30 min. The reaction mixture was evaporated to dryness togive4-(2,4-dichlorophenyl)-5-(3-(dimethylamino)acryloyl)-2-morpholinothiophene-3-carbonitrileas an intermediate. The above intermediate and hydrazine hydrate (30 mg,0.0006 mol) in AcOH (1.5 mL, 0.026 mol) was irradiated in microwave at120° C. for 15 min. The reaction mixture was evaporated and the residuewas purified by column chromatography to afford4-(2,4-dichlorophenyl)-2-morpholino-5-(1H-pyrazol-5-yl)thiophene-3-carbonitrile(19.5 mg, 84%). LCMS: (FA) ES+ 405.06, ES− 403.23; ¹H NMR (400 MHz,d₄-methanol) δ: 7.65 (d, 1H), 7.45 (m, 2H), 7.33 (d, 1H), 5.48 (s, 1H),5.45 (br, 1H), 3.85 (m, 4H), 3.54 (m, 4H).

Compounds in the following table were prepared from the appropriatestarting materials in a method analogous to that of Example 7:

29 LCMS: (FA) ES+ 419.

Example 8 Synthesis of4-(2,4-dichlorophenyl)-2-morpholino-5-(1H-tetrazol-5-yl)thiophene-3-carbonitrile(Compound I-23)

Step 1:4-(2,4-dichlorophenyl)-2-morpholino-5-(1H-tetrazol-5-yl)thiophene-3-carbonitrile

To a mixture of sodium azide (34.01 mg, 0.0005232 mol) in ACN (0.3 mL,0.005 mol) was added silicon(IV) chloride (0.105 mmol, 0.000105 mol) inDCM (0.1 mL, 0.002 mol) and the mixture was stirred for 30 min.4-cyano-3-(2,4-dichlorophenyl)-5-morpholinothiophene-2-carboxamide (20.0mg, 0.0000523 mol) was added to the above mixture, followed by sodiumazide (0.0340 g, 0.000523 mol) and silicon(IV) chloride (0.16 mmol,0.00016 mol) in DCM (0.16 mL, 0.0025 mol) and the mixture was irradiatedin microwave at 160° C. for 30 min. The reaction was evaporated andpurified by column chromatography to afford4-(2,4-dichlorophenyl)-2-morpholino-5-(1H-tetrazol-5-yl)thiophene-3-carbonitrile(11.6 mg, 54.4%). LCMS: (FA) ES+ 407.09, ES− 405.20; NMR (300 MHz,d₁-chloroform) δ: 7.60 (d, 1H), 7.42 (dd, 1H), 7.30 (d, 1H), 3.90-3.85(m, 4H), 3.72-3.63 (m, 4H).

Compounds in the following table were prepared from the appropriatestarting materials in a method analogous to that of Example 7:

6 LCMS: (FA) ES+ 389.22.

Example 9 Synthesis of4-(2,4-dichlorophenyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(Compound I-34) and4-(2,4-dichlorophenyl)-2-(tetrahydro-2H-pyran-4-yl)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(Compound I-5)

Step 1: Ethyl4-cyano-5-[(2,4-dimethoxybenzyl)amino]-3-iodothiophene-2-carboxylate

Ethyl 4-cyano-3-iodo-5-(methylsulfonyl)thiophene-2-carboxylate (5.60 g,0.0145 mol) and 2,4-dimethoxybenzylamine (3.51 mL, 0.0234 mol) werecombined in tetrahydrofuran (100 mL) and stirred at 60° C. for threedays. The reaction was concentrated in vacuo, diluted withdichloromethane and hexanes and the resultant precipitate was filteredto yield the title compound (5.56, 81%) as a yellow solid. LCMS: (FA)ES⁺, 473. ¹H NMR (400 MHz, d₆-DMSO) δ: 9.05 (s, 1H) 7.10 (d, 1H, J=8.57Hz), 6.60-6.50 (m, 2H), 4.30 (s, 2H), 4.22-4.14 (m, 2H), 3.80 (s, 3H),3.75 (s, 3H), 1.26-1.21 (m, 3H).

Step 2: Ethyl4-cyano-3-(2,4-dichlorophenyl)-5-[(2,4-dimethoxybenzyl)amino]thiophene-2-carboxylate

Ethyl4-cyano-5-[(2,4-dimethoxybenzyl)amino]-3-iodothiophene-2-carboxylate(3.18 g, 0.00673 mol) 2,4-dichlorophenylboronic acid (2.72 g, 0.0143mol), tetrakis(triphenylphosphine)palladium (0) (0.47 g, 0.00040 mol),and sodium carbonate (2.42 g, 0.0228 mol) were suspended in1,2-dimethoxyethane (23.5 mL) and water (13.5 mL). The suspension wasflushed with argon and the reaction mixture was irradiated in microwaveat 140° C. (300 watts) for ten minutes. The reaction mixture was dilutedwith a saturated solution of sodium bicarbonate in water and extractedwith ethyl acetate. The organic extracts were washed with brine, driedover anhydrous sodium sulfate, filtered and concentrated in vacuo.Column chromatography was performed to yield the title compound (2.92 g,88%). LCMS: (FA) ES⁺, 491. ¹H NMR (400 MHz, d₆-DMSO) δ: 9.05 (bs, 1H)7.75 (d, 1H, J=2.00 Hz), 7.52-7.48 (m, 1H), 7.40 (d, 1H, J=8.28 Hz),7.19 (d, 1H, J=8.53 Hz), 6.62-6.53 (m, 2H), 4.35 (bs, 2H), 4.04-3.92 (m,2H), 3.83 (s, 3H), 3.76 (s, 3H), 1.01-0.96 (m, 3H).

Step 3: Ethyl5-amino-4-cyano-3-(2,4-dichlorophenyl)thiophene-2-carboxylate

Ethyl4-cyano-3-(2,4-dichlorophenyl)-5-[(2,4-dimethoxybenzyl)amino]thiophene-2-carboxylate(4.70 g, 0.00956 mol) was dissolved in dichloromethane (100 mL).Trifluoroacetic acid (25 mL) was added and the solution was stirred atroom temperature for ten minutes. The reaction was concentrated invacuo, diluted with ethyl acetate and filtered. The filtrate was washedwith saturated sodium bicarbonate and brine, dried over anhydrous sodiumsulfate, filtered and concentrated in vacuo. Column chromatography wasperformed to yield the title compound (2.92 g, 90%) as a yellow solid.LCMS: (FA) ES⁺, 341. ¹H NMR (400 MHz, d₆-DMSO) δ: 8.17 (s, 2H) 7.75 (d,1H, J=2.00 Hz), 7.52-7.48 (m, 1H), 7.39 (d, 1H, J=8.28 Hz), 4.05-3.92(m, 2H), 1.02-0.96 (m, 3H).

Step 4: Ethyl4-cyano-3-(2,4-dichlorophenyl)-5-iodothiophene-2-carboxylate

To a suspension of ethyl5-amino-4-cyano-3-(2,4-dichlorophenyl)thiophene-2-carboxylate (2.92 g,0.00856 mol) in acetonitrile (10 mL) was added diiodomethane (2.41 mL,0.0300 mol) under an atmosphere of argon and was heated at 38° C.Isoamyl nitrite (2.61 g, 0.0214 mol) was added dropwise and the reactionmixture was cooled to room temperature and stirred for one hour. Thereaction was concentrated in vacuo and column chromatography wasperformed to yield the title compound (1.44 g, 37%) as an orange solid.¹H NMR (400 MHz, d₁-chloroform) δ: 7.53 (d, 1H, J=2.00 Hz), 7.38-7.34(m, 1H), 7.21 (d, 1H, J=8.28 Hz), 4.25-4.15 (m, 2H), 1.21-1.16 (m, 3H).

Step 5: 4-cyano-3-(2,4-dichlorophenyl)-5-iodothiophene-2-carboxylic acid

To a solution of ethyl4-cyano-3-(2,4-dichlorophenyl)-5-iodothiophene-2-carboxylate (1.44 g,0.00318 mol) in tetrahydrofuran (20 mL) and water (10 mL) was added asolution of 1.00M sodium hydroxide in water (16 mL). The solution wasallowed to stir overnight. The reaction was quenched with a solution of1N hydrogen chloride in water (18 mL) and extracted with ethyl acetate.The organic extracts were washed with brine, dried over anhydrous sodiumsulfate, filtered and concentrated in vacuo to afford the crude titlecompound (1.50 g, 100%) used directly in the next reaction. LCMS: (FA)ES⁺, 378. ¹H NMR (400 MHz, d₆-DMSO) δ: 7.68 (d, 1H, J=2.0 Hz), 7.46-7.34(m, 2H).

Step 6: 4-cyano-3-(2,4-dichlorophenyl)-5-iodothiophene-2-carboxylamide

4-cyano-3-(2,4-dichlorophenyl)-5-iodothiophene-2-carboxylic acid (1.30g, 0.00306 mol) was dissolved in dichloromethane (30 mL).N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.27 g,0.00661 mol) and 1-hydroxybenzotriazole (0.880 g, 0.00651 mol) wereadded to the solution and the reaction was stirred for thirty minutes.Ammonium hydroxide (5.97 mL, 0.153 mol) was added to the solution andthe biphasic mixture was stirred for two hours. The reaction mixture wasconcentrated, diluted with water and extracted with ethyl acetate. Theorganic extract was dried over anhydrous magnesium sulfate, filtered andcolumn chromatography was performed to yield the title compound (1.21 g,89%). LCMS: (FA) ES⁺, 423. ¹H NMR (400 MHz, d₆-DMSO) δ: 7.79 (d, 1H,J=2.0 Hz), 7.68 (bs, 1H), 7.57-7.45 (m, 2H), 7.30 (bs, 1H).

Step 7:4-(2,4-dichlorophenyl)-2-iodo-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile

A mixture of4-cyano-3-(2,4-dichlorophenyl)-5-iodothiophene-2-carboxylamide (1.33 g,0.00314 mol) and 1,1-dimethoxy-N,N-dimethylmethanamine (10.0 mL, 0.0753mol) was irradiated in the microwave at 120° C. (300 watts) for 30minutes. The reaction was concentrated in vacuo. The residue dissolvedin acetic acid (1.0 mL, 0.18 mol) and hydrazine hydrate (0.69 mL, 0.014mol) and subjected to microwave irradiation at 120° C. (300 watts) for15 minutes. The solvent was removed in vacuo and the residue wasazeotroped with toluene. Column chromatography was performed to yieldthe title compound (1.25 g, 85%). LCMS: (FA) ES⁺, 447. ¹H NMR (400 MHz,d₄-methanol) δ: 8.35 (s, 1H) 7.60 (d, 1H, J=2.0 Hz), 7.45-7.35 (m, 2H).

Step 8:4-(2,4-dichlorophenyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile

A mixture of4-(2,4-dichlorophenyl)-2-iodo-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(180 mg, 0.4 mmol), tributyl(3,6-dihydro-2H-pyran-4-yl)stannane (0.452g, 1.2 mmol), lithium chloride (51.2 mg, 1.2 mmol), copper(I) iodide(7.6 mg, 0.04 mmol), tetrakis(triphenylphosphine)palladium (46.4 mg,0.04 mmol) was dissolved in dioxane (20 mL) and heated to reflux for 3hours under an atmosphere of argon. The solvent was removed and theresidue was purified using ISCO chromatography on silica gel, elution20% ethyl acetate in hexanes to ethyl acetate to afford the titlecompound (24 mg, 14%). LCMS: (FA) ES⁺, 403. ¹H NMR (400 MHz,d₄-methanol) δ: 8.35 (s, 1H), 7.59 (d, 1H), 7.43-7.36 (m, 2H), 6.72 (dd,1H), 4.38-4.36 (m, 2H), 3.97-3.94 (m, 2H), 2.71-2.68 (m, 2H).

Step 9:4-(2,4-dichlorophenyl)-2-(tetrahydro-2H-pyran-4-yl)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile

4-(2,4-dichlorophenyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-(4H-1,2,4-triazol-3-yl)thiophene-3-carbonitrile(20 mg, 0.05 mmol) was dissolved in methanol (2 mL) and 10% Pd/C wasadded (10 mg). The mixture was stirred under an atmosphere of hydrogenfor 2 hours. The suspension was filtered through celite, solvent wasevaporated and the residue was purified using ISCO chromatography onsilica gel, elution 20% ethyl acetate in hexanes to ethyl acetate toafford the title compound (16 mg, 80%). LCMS: (FA) ES⁺, 405. ¹H NMR (400MHz, d₄-methanol) δ: 8.53 (s, 1H), 7.78 (d, 1H), 7.51-7.46 (m, 2H),4.12-4.08 (m, 1H), 3.99-3.95 (m, 1H), 3.53-3.30 (m, 2H), 3.20-3.15 (m,1H), 2.00-1.95 (m, 1H), 1.83-1.72 (m, 1H).

Example 10 Synthesis of4-[4-(2,4-dichlorophenyl)-5-(4H-1,2,4-triazol-3-yl)-1,3-thiazol-2-yl]morpholine(Compound I-1)

Step 1: Synthesis of4-chloro-2-morpholin-4-yl-1,3-thiazole-5-carbonitrile

2,4-dichloro-5-cyanothiazole (1.26 g, 0.00704 mol) andN,N-diisopropylethylamine (3.68 mL, 0.0211 mol) were dissolved inethanol at 70° C. Morpholine (0.614 mL, 0.0704 mol) was added to the hotsolution and the mixture was stirred for 30 minutes, cooled down to rt,diluted with water (50 mL) and the formed precipitate was collected togive the title compound (1.55 g, 91%). LCMS: (FA) ES⁺, 230. ¹H NMR (300MHz, d₆-DMSO) δ: 3.71-3.67 (m, 4H), 3.52-3.48 (m, 4H).

Step 2: Synthesis of4-(2,4-dichlorophenyl)-2-morpholin-4-yl-1,3-thiazole-5-carbonitrile

4-chloro-2-morpholin-4-yl-1,3-thiazole-5-carbonitrile (0.200 g, 0.871mmol), 2,4-dichlorophenylboronic acid (0.249 g, 1.31 mmol),bis(triphenylphosphine)palladium dichloride (0.061 g, 0.0871 mmol),sodium carbonate (0.184 g, 1.74 mmol) were taken up in DME (3 mL),ethanol (1 mL) and water (1 mL) and microwaved at 125° for 30 minutes.Mixture was diluted with ethyl acetate (10 mL) and extracted. Theorganic phase was dried with sodium sulfate, filtered and evaporated.ISCO purification on silica gel using 10% ethyl acetate in hexane to 50%ethyl acetate in hexane afforded the title compound (45 mg, 15%). LCMS:(FA) ES⁺, 340. ¹H NMR (400 MHz, d₆-DMSO) δ: 7.44 (d, 1H), 7.20-7.17 (m,2H), 3.43-3.28 (m, 4H), 3.16-2.93 (m, 4H).

Step 3: Synthesis of4-(2,4-dichlorophenyl)-2-morpholin-4-yl-1,3-thiazole-5-carboxamide

4-(2,4-dichlorophenyl)-2-morpholin-4-yl-1,3-thiazole-5-carbonitrile (30mg, 0.088 mmol) was dissolved in sulfuric acid (0.200 mL, 3.75 mmol) andthe mixture was stirred at rt for 1 hour. Water (1 mL) was added and themixture was quenched with an excess of saturated NaHCO₃. Precipitate wasfiltered off, washed with water and dried to afford the title compound(27 mg, 80%). LCMS: (FA) ES⁺, 358. ¹H NMR (400 MHz, d₆-DMSO) δ: 7.67 (d,1H), 7.47 (d, 1H), 7.43 (dd, 1H), 3.72-3.69 (m, 4H), 3.44-3.42 (m, 4H).

Step 4: Synthesis of4-[4-(2,4-dichlorophenyl)-5-(4H-1,2,4-triazol-3-yl)-1,3-thiazol-2-yl]morpholine

A mixture of4-(2,4-dichlorophenyl)-2-morpholin-4-yl-1,3-thiazole-5-carboxamide (23mg, 0.06 mmol) and 1,1-dimethoxy-N,N-dimethylmethanamine (0.8 mL, 6 mol)was irradiated in the microwave at 160° C. (300 watts) for 30 minutes.The reaction was concentrated in vacuo. The residue dissolved in aceticacid (1.0 mL, 0.18 mol) and hydrazine hydrate (30 mg, 0.7 mmol) andsubjected to microwave irradiation at 120° C. (300 watts) for 15minutes. The solvent was removed in vacuo, the residue was diluted withwater (1 mL), the precipitate was collected, washed with water (1 mL)and dried to afford the title compound (10 mg, 40%). LCMS: (FA) ES⁺,382. ¹H NMR (400 MHz, d₆-DMSO) δ: 13.9 (s, 1H), 8.42 (s, 1H), 7.63 (d,1H), 7.44-7.42 (m, 2H), 3.73-3.70 (m, 4H), 3.44-3.42 (m, 4H).

II. Biological Data Example 1 PI3K Enzyme Assay

Expression and Purification of PI3K Enzyme

Active phosphatidylinositol 3′ kinase (PI3K) enzyme was purified atMillennium Pharmaceuticals from SF9 insect cells (Invitrogen)co-infected with baculovirus containing amino-terminal His-tagged p110αand p85α expression constructs.

PI3K Enzyme Homogenous Time Resolved Fluorescence (HTRF®) Assay

The PI3K enzyme HTRF® assay makes use of an energy transfer complexcomprised of biotin-PI(3,4,5)P₃, Europhium labeled anti-GST monoclonalantibody, a GST-tagged GRP1 pleckstrin homology (PH) domain, andStreptavidin-APC (allophycocyanin). Excitation of the Europium in thecomplex results in a stable time-resolved fluorescence resonance energytransfer (FRET) signal. Phosphatidylinositol 3,4,5 triphosphate(PI(3,4,5)P₃, the product of PI3K, disrupts the energy transfer complexby competing with biotin-PI(3,4,5)P₃ for binding to the GRP1 PH domain,resulting in a decreased fluorescent signal. Inhibitors of PI3K in thereaction prevent a decrease in the fluorescent signal.

PI3K enzyme (325 pM) was incubated with di-C8 PI(4,5)P₂ substrate (3.5μM, CellSignals, Inc.) in assay buffer (50 mM HEPES pH 7.0, 5 mM DTT,150 mM NaCl, 10 mM β-glycerophosphate, 5 mM MgCl₂, 0.25 mM sodiumcholate, 0.001% CHAPS) containing 25 μM ATP and 0.5 μL of test compound(in 100% DMSO) at multiple concentrations in a final volume of 20.5 μLin 384 well plates for 30 min at 22-23° C. The reaction was terminatedby adding 5 μL of detection buffer (50 mM HEPES pH7.0, 5 mM DTT, 1 mMNaCl, 10% Tween-20) containing EDTA (90 mM) and biotin-PI(3,4,5)P₃ (150nM, Echelon Bioscience) to each well. 5 μL of detection buffercontaining GST-fused GRP1 PH domain protein (210 nM, MillenniumPharmaceuticals), anti-GST-Europium tagged cryptate antibody (2.25 nM,CisBio), Streptavidin-XL (90 nM, CisBio) and potassium fluoride (240 mM)were then added to each well and incubated for 1 hour. Fluorescentsignal for each well was then measured on an LJL_Analyst (MolecularDevices). Concentration response curves were generated by calculatingthe fluorescent signal in test compound-treated samples relative toDMSO-treated (0% inhibition) and EDTA-treated (100% inhibition)controls, and concentrations producing 50% inhibition (IC₅₀ values) weredetermined from those curves.

Example 2 PI3K Cell Assays

Forkhead Redistribution Assay

Inhibition of PI3K in cells can be assessed using the ForkheadRedistribution Assay (BioImage). Foxo1A fused to EGFP (Foxo1A-EGFP)expressed in U2OS osteosarcoma cells localizes to the cytoplasm when thePI3K pathway is actively signaling. Inactivation of pathway signalingleads to a translocation of the protein from the cytoplasm to thenucleus. Therefore, pathway inhibition can be measured by quantifyingthe fluorescent intensity of Foxo1A-EGFP within the nucleus.

U2OS cells constitutively expressing Foxo1A-EGFP (6500 cells/well) wereplated onto the inner 60 wells of 96 well dishes (BD Falcon OPTILUXblack clear bottom) in 100 μL of cell culture media (DMEM (Invitrogen)containing 10% Fetal Bovine Serum (HyClone) and 1%Penicillin-Streptavidin (Invitrogen) and grown overnight in a humidifiedchamber at 37° C. The cell culture media was removed and the cells wererinsed with 100 μL of low serum media (DMEM containing 0.933% FetalBovine Serum and 1% Penicillin-Streptavadin) and incubated in 75 μL oflow serum media for 1 hour in a humidified chamber at 37° C. Testcompounds (25 μL) at multiple concentrations suspended in DMEMcontaining 1% Penicillin-Streptavadin were added to cells and incubatedin a humidified chamber at 37° C. for 1 hour. The media was removed andthe cells were fixed in 100 μL of 4% paraformaldehyde in phosphatebuffered saline (PBS) for 10 min and then washed with 100 μL of PBS.DRAQ5 mix (100 μL, Alexis Biochemicals) diluted 1:5000 in PBS containingRNAase (1:10,000, Sigma) was added to cells for 30 minutes. The plateswere then imaged (16 fields per well) using an Opera Imager (Evotec) andFoxo1A-EGFP fluorescent intensity within the nucleus (DRAQ5-positive)was quantified using Acapella Software (Evotec). Concentration responsecurves were generated by calculating the nuclear fluorescent intensityof Foxo-1A EGFP in test compound-treated samples and concentrationsproducing 50% inhibition (IC₅₀ values) relative to the positive controlwere determined from those curves.

Example 3 Anti-Proliferation Assay

ATPlite Assay

The ATPLite™ (Perkin-Elmer) Assay measures cellularadenosine-triphosphate (ATP) through the generation of a luminescentsignal formed from the ATP dependent enzyme firefly luciferase. Theluminescent signal intensity can be used as a measure of cellularproliferation, and therefore the anti-proliferative effects of PI3Kinhibitors.

Test compounds (4 μL in 100% DMSO) were diluted in 75 μL of HanksBuffered Saline Solution (Invitrogen). The diluted test compounds (8 μL)were then added to 384-well TC-treated Black/Clear plates (Falcon).HCT-116 cells (American Type Culture Collection) maintained in McCoy's5a modified media (Invitrogen) containing 10% Fetal Bovine Serum and 1%Penicillin-Streptavadin were added at 1000 cells per well. H460 cells(American Type Culture Collection) maintained in RPMI 1640 containing10% Fetal Bovine Serum and 1% Penicillin-Streptavadin were added at 1500cells per well. The cells were then incubated with compound in ahumidified chamber at 37° C. for 72 hours. The plates were then removedfrom the cell culture chambers and allowed to equilibrate to roomtemperature for 30 min. All but 25 μL of cell culture media was removedfrom each well, and 25 μl of ATPlite reagent (Perkin Elmer) was added toeach well. Luminescence was measured within 5 minutes of adding theATPlite reagent on a LEADSeeker Luminescence Counter (GE Healthcare LifeSciences). Concentration response curves were generated by calculatingthe luminescence decrease in test compound-treated samples relative toDMSO-treated controls, and growth inhibition (IC₅₀) values weredetermined from those curves.

As detailed above, compounds of the invention inhibit PI3K. In certainembodiments, compounds of the invention have an IC50<5.0 μM. In otherembodiments, compounds of the invention have an IC50<1.0 μM. In stillother embodiments, compounds of the invention have an IC50<0.1 μM.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments, which utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments, which have been represented by way of example.

As detailed above, compounds of the invention inhibit PI3K. In certainembodiments, compounds of the invention have an IC50<5.0 μM. In otherembodiments, compounds of the invention have an IC50<1.0 μM. In stillother embodiments, compounds of the invention have an IC50<0.1 μM.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments, which utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments, which have been represented by way of example.

1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: when Y₂ is C, R¹ is H, —CN, halogen, —Z—R³, C₁₋₆ aliphatic, or 3-10-membered cycloaliphatic, wherein: Z is selected from an optionally substituted C₁₋₃ alkylene chain, —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —CO₂—, —C(O)NR^(1a)—, —N(R^(1a))C(O)—, —N(R^(1a))CO₂—, —S(O)₂NR^(1a)—, —N(R^(1a))S(O)₂—, —OC(O)N(R^(1a))—, —N(R^(1a))C(O)NR^(1a)—, —N(R^(1a))S(O)₂N(R^(1a))—, or —OC(O)—; R^(1a) is hydrogen or an optionally substituted C₁₋₄ aliphatic, and R³ is an optionally substituted group selected from C₁₋₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; R² is H, halogen, —W—R⁵, or —R⁵, wherein: W is selected from an optionally substituted C₁₋₃ alkylene chain, —O—, —N(R^(2a))—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —CO₂—, —C(O)NR^(2a)—, —N(R^(2a))C(O)—, —N(R^(2a))CO₂—, —S(O)₂NR^(2a)—, N(R^(2a))S(O)₂—, —OC(O)N(R^(2a))—, —N(R^(2a))C(O)NR^(2a)—, —N(R^(2a))S(O)₂N(R^(2a))—, or —OC(O)—. R^(2a) is hydrogen or an optionally substituted C₁₋₄ aliphatic, and R⁵ is an optionally substituted group selected from C₁₋₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; X₁, X₂, and X₃ are each independently N or CR⁶, wherein each occurrence of R⁶ is independently hydrogen, —CN, halogen, —V—R⁷, C₁₋₆ aliphatic, or 3-10-membered cycloaliphatic, wherein: V is selected from an optionally substituted C₁₋₃ alkylene chain, —O—, —N(R^(6a))—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —CO₂—, —C(O)NR^(6a)—, —N(R^(6a))C(O)—, —N(R^(6a))CO₂—, —S(O)₂NR^(6a)—, —N(R^(6a))S(O)₂—, —OC(O)N(R^(6a))—, —N(R^(6a))C(O)NR^(6a)—, —N(R^(6a))S(O)₂N(R^(6a))—, or —OC(O)—. R^(6a) is hydrogen or an optionally substituted C₁₋₄ aliphatic, and R⁷ is an optionally substituted group selected from C₁₋₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; Y₁ is S, O, NR⁸, wherein R⁸ is hydrogen or an optionally substituted C₁₋₄aliphatic; Y₂ is C, N CY is

wherein each occurrence of R⁴ is independently-R^(4a) or -T₁-R^(4d), wherein: each occurrence of R^(4a), as valency and stability permit, is independently fluorine, ═O, ═S, —CN, —NO₂, —R^(4c), —N(R^(4b))₂, —OR^(4b), —SR^(4c), —S(O)₂R^(4c), —C(O)R^(4b), —C(O)OR^(4b), —C(O)N(R^(4b))₂, —S(O)₂N(R^(4b))₂, —OC(O)N(R^(4b))₂, —N(R^(4e))C(O)R^(4b), —N(R^(4e))SO₂R^(4c), —N(R^(4e))C(O)OR^(4b), —N(R^(4e))C(O)N(R^(4b))₂, or —N(R^(4e))SO₂N(R^(4b))₂, or two occurrences of R^(4b), taken together with a nitrogen atom to which they are bound, form an optionally substituted 4-7-membered heterocyclyl ring having 0-1 additional heteroatoms selected from nitrogen, oxygen, or sulfur; each occurrence of R^(4b) is independently hydrogen or an optionally substituted group selected from C₁-C₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(4c) is independently an optionally substituted group selected from C₁-C₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(4d) is independently hydrogen or an optionally substituted from 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(4e) is independently hydrogen or an optionally substituted C₁₋₆ aliphatic group; and T₁ is an optionally substituted C₁-C₆alkylene chain wherein the alkylene chain optionally is interrupted by —N(R^(4a))—, —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(4a))—, —S(O)₂N(R^(4a))—, —OC(O)N(R^(4a))—, —N(R^(4a))C(O)—, —N(R^(4a))SO₂—, —N(R^(4a))C(O)O—, —NR^(4a)C(O)N(R^(4a))—, —N(R^(4a))S(O)₂N(R^(4a))—, —OC(O)—, or —C(O)N(R^(4a))—O— or wherein T₁ or a portion thereof optionally forms part of an optionally substituted 3-7 membered cycloaliphatic or heterocyclyl ring; n is 0-6; m is 1 or 2; p is 0, 1, or 2;

represents a single or double bond; and provided that the compound of formula I is other than Morpholine, 4-[5-(4,5-diphenyl-1H-imidazol-2-yl)-2-thienyl]-.
 2. The compound of claim 1, wherein one or more substituents are selected from: (a) Y₁ is S; (b) Y₂ is C (c) R¹ is CN or H; (d) R² is an optionally substituted 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; (e) n is 0-2; or (f) R⁴ is —R^(4a).
 3. The compound of claim 2, wherein the compound is represented by:


4. The compound of claim 1, wherein R² is a 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, optionally substituted with 1-4 independent occurrences of R⁹, wherein R⁹ is —R^(9a), -T₂-R^(9d), or —V₂-T₂-R^(9d), and: each occurrence of R^(9a) is independently halogen, —CN, —NO₂, —R^(9c), —N(R^(9b))₂, —OR^(9b), —SR^(9c), —S(O)₂R^(9c), —C(O)R^(9b), —C(O)OR^(9b), —C(O)N(R^(9b))₂, —S(O)₂N(R^(9b))₂, —OC(O)N(R^(9b))₂, —N(R^(9e))C(O)R^(9b), —N(R^(9e))SO₂R^(9c), —N(R^(9e))C(O)OR^(9b), —N(R^(9e))C(O)N(R^(9b))₂, or —N(R^(9e))SO₂N(R^(9b))₂, or two occurrences of R^(9b), taken together with a nitrogen atom to which they are bound, form an optionally substituted 4-7-membered heterocyclyl ring having 0-1 additional heteroatoms selected from nitrogen, oxygen, or sulfur; each occurrence of R^(9b) is independently hydrogen or an optionally substituted group selected from C₁-C₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(9c) is independently an optionally substituted group selected from C₁-C₆aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(9d) is independently hydrogen or an optionally substituted from 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(9e) is independently hydrogen or an optionally substituted C₁₋₆ aliphatic group; each occurrence of V₂ is independently —N(R^(9e))—, —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(9e))—, —S(O)₂N(R^(9e))—, —OC(O)N(R^(9e))—, —N(R^(9e))C(O)—, —N(R^(9e))SO₂—, —N(R^(9e))C(O)O—, —NR^(9e)C(O)N(R^(9e))—, —N(R^(9e))SO₂N(R^(9e))—, —OC(O)—, or —C(O)N(R^(9e))—O—; and T₂ is an optionally substituted C₁-C₆ alkylene chain wherein the alkylene chain optionally is interrupted by —N(R^(7a))—, —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(7a))—, —S(O)₂N(R^(7a))—, —OC(O)N(R^(7a))—, —N(R^(7a))C(O)—, —N(R^(7a))SO₂—, —N(R^(7a))C(O)O—, —NR^(7a)C(O)N(R^(7a))—, —N(R^(7a))S(O)₂N(R^(7a))—, —OC(O)—, or —C(O)N(R^(7a))—O— or wherein T₃ or a portion thereof optionally forms part of an optionally substituted 3-7 membered cycloaliphatic or heterocyclyl ring.
 5. The compound of claim 1, wherein Y₁ is S, Y₂ is C and the compound is represented by formula II:

wherein R¹ is CN or H.
 6. The compound of claim 5, wherein: R² is an optionally substituted 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; n is 0-2; and R⁴ is —R^(4a).
 7. The compound of claim 6, wherein the compound is represented by:


8. The compound of claim 7, wherein: R² is an optionally substituted 6-10-membered aryl, or a 5-10-membered heteroaryl ring having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; wherein R² is optionally substituted with 1-4 independent occurrences of R⁹, wherein R⁹ is —R^(9a), -T₂-R^(9d), or —V₂-T₂-R^(9d), and: each occurrence of R^(7a) is independently halogen, —CN, —NO₂, —R^(7c), —N(R^(7b))₂, —OR^(7b), —SR^(7c), —S(O)₂R^(9c), —C(O)R^(9b), —C(O)OR^(9b), —C(O)N(R^(9b))₂, —S(O)₂N(R^(9b))₂, —OC(O)N(R^(9b))₂, —N(R^(9e))C(O)R^(9b), —N(R^(9e))SO₂R^(9c), —N(R^(9e))C(O)OR^(9b), —N(R^(9e))C(O)N(R^(9b))₂, or —N(R^(9e))SO₂N(R^(9b))₂; each occurrence of R^(9b) is independently hydrogen or an optionally substituted group selected from C₁-C₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or two occurrences of R^(9b), taken together with a nitrogen atom to which they are bound, form an optionally substituted 4-7-membered heterocyclyl ring having 0-1 additional heteroatoms selected from nitrogen, oxygen, or sulfur; each occurrence of R⁹ is independently an optionally substituted group selected from C₁-C₆ aliphatic, 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(9d) is independently hydrogen or an optionally substituted from 3-10-membered cycloaliphatic, 4-10-membered heterocyclyl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 6-10-membered aryl, or 5-10-membered heteroaryl having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each occurrence of R^(9e) is independently hydrogen or an optionally substituted C₁₋₆ aliphatic group; each occurrence of V₂ is independently —N(R^(7e))—, —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(9e))—, —S(O)₂N(R^(9e))—, —OC(O)N(R^(9e))—, —N(R^(9e))C(O)—, —N(R^(9e))SO₂—, —N(R^(9e))C(O)O—, —NR^(9e)C(O)N(R^(9e))—, —N(R^(9e))SO₂N(R^(9e))—, —OC(O)—, or —C(O)N(R^(9e))—O—; and T₂ is an optionally substituted C₁-C₆alkylene chain wherein the alkylene chain optionally is interrupted by —N(R^(9a))—, —O—, —S—, —S(O)—, —S(O)₂—, —C(O)—, —C(O)O—, —C(O)N(R^(9a))—, —S(O)₂N(R^(9a))—, —OC(O)N(R^(9a))—, —N(R^(9a))C(O)—, —N(R^(9a))SO₂—, —N(R^(9a))C(O)O—, —NR^(9a)C(O)N(R^(9a))—, —N(R^(9a))S(O)₂N(R^(9a))—, —OC(O)—, or —C(O)N(R^(9a))—O— or wherein T₂ or a portion thereof optionally forms part of an optionally substituted 3-7 membered cycloaliphatic or heterocyclyl ring; and n is 0-2.
 9. The compound of claim 8, wherein: R² is a phenyl group substituted with 1-3 independent occurrences of halogen, —CN, —NO₂, —R^(9c), —N(R^(9b))₂, —OR^(9b), —SR^(9c), —S(O)₂R^(9c), —C(O)R^(9b), —C(O)OR^(9b), —C(O)N(R^(9b))₂, —S(O)₂N(R^(9b))₂, —OC(O)N(R^(9b))₂, —N(R^(9e))C(O)R^(9b), —N(R^(9e))SO₂R^(9c), —N(R^(9e))C(O)OR^(9b), —N(R^(9e))C(O)N(R^(9b))₂, or —N(R^(9e))SO₂N(R^(9b))₂; R³ is hydrogen, C₁₋₄alkyl, or C₁₋₄fluoroalkyl; and n is
 0. 10. The compound of claim 9, wherein: R² is a phenyl group substituted with 1-3 independent occurrences of halo, C₁₋₃ alkyl, CN, C₁₋₃haloalkyl, —OC₁₋₃ alkyl, —OC₁₋₃ haloalkyl, —NHC(O)C₁₋₃ alkyl, —NHC(O)NHC₁₋₃ alkyl, NHS(O)₂C₁₋₃ alkyl, or —COH.
 11. The compound of claim 1, wherein the compound is selected from:


12. A composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier.
 13. A method of treating a proliferative disorder in a patient comprising administering to said patient a therapeutically effective amount of a compound of claim
 1. 14. The method of claim 13, wherein the proliferative disorder is breast cancer, bladder cancer, colon cancer, glioma, glioblastoma, lung cancer, hepatocellular cancer, gastric cancer, melanoma, thyroid cancer, endometrial cancer, renal cancer, cervical cancer, pancreatic cancer, esophageal cancer, prostate cancer, brain cancer, or ovarian cancer.
 15. A method of treating an inflammatory or cardiovascular disorder in a patient comprising administering to said patient a therapeutically effective amount of a compound of claim
 1. 16. The method of claim 15, wherein the inflammatory or cardiovascular disorder is selected from allergies/anaphylaxis, acute and chronic inflammation, rheumatoid arthritis; autoimmunity disorders, thrombosis, hypertension, cardiac hypertrophy, and heart failure.
 17. A method for inhibiting PI3K activity in a patient comprising administering a composition comprising an amount of a compound of claim 1 effective to inhibit PI3K activity in the patient. 